WO2016031705A1 - Magnetic particle reaction control apparatus utilizing variable-pitch dispensing apparatus and reaction control method therefor - Google Patents

Abstract

The present invention provides a magnetic particle reaction control apparatus utilizing a variable-pitch dispensing apparatus, which prevents expansion of apparatus scale and is capable of handling containers of various pitches, and a reaction control method therefor. The invention is provided with a nozzle head having a plurality of nozzles which have tip sections and are arranged in a single row at a variable pitch, and a pitch conversion mechanism capable of converting the nozzles between a first pitch and a second pitch by command; a first receiving section group having receiving sections which are arranged, at a first pitch, in at least one row along a row direction, and into which the tip sections can be inserted simultaneously, and a second receiving section group having receiving sections which are arranged, at a second pitch smaller than the first pitch, in at least one row in the row direction; a movement mechanism which allows the nozzle head or nozzles to be moved with respect to the receiving section groups; and a magnetic device capable of applying and removing a magnetic field, simultaneously inside the tip sections of the nozzles arranged at the first pitch or the second pitch; wherein either the first receiving section group or the second receiving section group contains magnetic particle suspensions.

Description

Magnetic particle reaction control device using variable pitch dispensing device and reaction control method thereof

The present invention relates to a magnetic particle reaction control device using a variable pitch dispensing device and a reaction control method thereof.

Conventionally, for a plurality of specimens such as whole blood collected from a plurality of patients, in order to perform various tests in parallel, for each specimen, collect a quantity of specimens according to the number of examinations, It must be stored in a specimen sample container such as a blood collection tube, and a plurality of types of reagents corresponding to the examination type must be prepared and dispensed and arranged according to the examination order. . For this purpose, for example, a nozzle having a number of nozzles equal to the number of specimens after the collected specimens are once stored in a plurality of containers such as test tubes and dispensed in advance into wells of a microplate. Processing is performed by sequentially moving the head along the row direction or the column direction with respect to the microplate (Patent Document 1).

The microplate is provided with a plurality of wells that can store liquids arranged in a matrix (matrix), for example, 4 rows × 6 columns (= 24 wells), 6 rows × 8 columns (= 48 wells), 8 rows × 12 columns (= 96 wells), 12 rows × 16 columns (= 192 wells), 16 rows × 24 columns (= 384 wells) are known. The pitch (distance between centers) of each well of these microplates tends to be standardized. For example, in a 96-well microplate, the pitch in the row direction and the column direction is 9 mm each, and the number of wells is When the number is doubled, the pitches are 4.5 mm and 9 mm, respectively, and when the number of wells is quadrupled, each pitch is 4.5 mm.

By the way, when a test requiring various reagents is performed on a large number of samples (for example, 8 samples), a microplate having a large number of wells (for example, 96 wells) is used. And the opening or capacity of each well is reduced. On the other hand, a certain amount of the sample is collected in order to cope with a plurality of examinations. In addition, in order to use each reagent for a plurality of specimens, it is necessary to prepare a certain amount in the reagent bottle. For example, when the specimen is whole blood, a tube with a diameter of 16 mm is usually used as a blood collection tube, and the pitch is, for example, 25 mm. Further, as the reagent bottle, for example, a wide-mouth bottle with a diameter of 25 mm is used, and the pitch thereof is, for example, 30 mm.

In that case, in the processing using the dispensing device, a single mounting nozzle with a dispensing tip attached to one of a plurality of specimen sample containers such as the blood collection tube is moved, and the dispensing is performed. The sample such as the whole blood is aspirated through the injection tip, moved to a predetermined well of one or two or more microplates, discharged or dispensed by a predetermined amount, and then the dispensing tip is detached from the nozzle. This is repeated for a plurality of blood collection tubes. As a result, the specimens contained in a plurality of specimen specimen containers are arranged on a microplate, and a new dispensing tip is attached to the attachment nozzle, and the reagent contained in one or more reagent bottles is removed. Dispensing into predetermined wells of the microplate and moving a nozzle head having a plurality of separately prepared nozzles equipped with a plurality of dispensing tips to process a plurality of specimens in parallel (Patent Document 2).

At that time, a magnet is provided in the nozzle head, a magnetic field is applied from the outside to the inside of the dispensing tip, and the magnetic particles that hold the target substance are adsorbed on the inner wall of the dispensing tip, and the magnetic field is removed. Then, the magnetic particles are resuspended in the liquid to transfer the magnetic particles between the wells (Patent Documents 1 and 2).

Also, for example, prepare multiple dispensing units with variable-pitch nozzle heads with 8 nozzles, and move the microplate between dispensing units manually or by container transfer means such as a conveyor. There was something to process. In this method, first, the microplate is positioned on the specimen dispensing unit device, the variable pitch of the nozzle head is set to 25 mm, moved to the blood collection tube, the whole blood is sucked, and then the variable pitch is set. Set to 9 mm, the nozzle head is moved to the microplate, and the specimen is sequentially ejected and dispensed at locations according to the number of examinations (specimen dispensing process). Next, the microplate is moved to the reagent dispensing unit device manually or by container transfer means such as a conveyor, and the variable pitch is set to 30 mm using a variable pitch having, for example, eight nozzles. After moving to the reagent bottle and aspirating the reagent, set the variable pitch to 9 mm, move the nozzle head to the microplate, and place the reagent at the appropriate location according to the test contents. Discharge and dispense sequentially (reagent dispensing step). Next, detection or measurement was performed by moving the microplate to a detection / measurement unit device by manual operation or container transfer means such as a conveyor (Patent Documents 3 and 4).

This is because a nozzle head with a variable pitch has a complicated nozzle head structure, and it is difficult to provide a magnetic device with a more complicated structure on the nozzle head, even if magnetic particles are used. The magnetic device applies a magnetic field to the container so that the magnetic particles are adsorbed on the container, and only the liquid to be treated is introduced into the dispensing tip provided in the nozzle head and transferred between the containers. Alternatively, the target substance is transferred.

As described above, conventionally, a nozzle head composed of a plurality of nozzles and magnets having a fixed pitch is used while transferring between containers having different pitches using a nozzle head composed of a single nozzle. A plurality of dedicated unit devices using nozzle heads with variable pitch are prepared separately for nozzles and magnets, and containers are manually or transferred to each unit device. It is transported by a mechanism. In the former case, in addition to the moving mechanism that moves a single nozzle, it is necessary to provide a moving mechanism that moves a plurality of nozzles with a fixed pitch. In the latter case, the nozzle also has a variable pitch. It is necessary to prepare a plurality of dedicated unit devices using the head separately for the nozzle and the magnet, which complicates the device structure, increases the manufacturing cost, makes the processing complicated, and increases the burden on the user. At the same time, there has been a problem that the scale of the apparatus is increased or there is a risk that a mistake in the specimen will occur (Patent Documents 1 to 4).

Furthermore, as the number of parallel processes handled in parallel increases, the number of wells in the microplate increases and the need for integration increases, and it is necessary to provide each nozzle closely, so the spacing between dispensing tips is increased. The problem is that it becomes increasingly difficult to add other functions such as complex magnetic devices around each dispensing tip, and it may become more difficult to handle magnetic particles as the pitch decreases. Had. In particular, in order to perform real-time PCR, chemiluminescence immunoassay, etc., light measurement and temperature control are required, and since the scale of the apparatus is increased, the necessity for integration of microplates becomes particularly high. In the case of performing a consistent treatment together with nucleic acid extraction, there is a problem in that there are many kinds of pitches of containers used, and it may become more difficult to handle magnetic particles.

Here, “real-time PCR” refers to a method of monitoring a nucleic acid (DNA) amplified by PCR in real time using a fluorescent substance. Real-time PCR has the advantage that amplification can be observed during the temperature cycle and that quantitative results can be obtained. Examples of methods usually performed using a fluorescent reagent containing a fluorescent substance include an intercalation method, a hybridization method, and a LUX method (Patent Document 5).

International Publication No. WO99 / 47267 Japanese Patent No. 3115501 JP 2007-085885 A JP 2004-340906 A International Publication No. WO2011 / 016509

Accordingly, the present invention has been made to solve the above-described problems, and a first object thereof is to have accommodating portions such as wells arranged at various pitches without increasing the scale of the apparatus. Magnetism using a variable pitch dispenser suitable for reaction processing using magnetic particles (for example, DNA extraction, target DNA isolation, real-time PCR pretreatment, immunoassay) for a container group such as a microplate An object of the present invention is to provide a particle reaction control device and a reaction control method thereof.

The second objective is suitable for automation that can perform from sample collection to sample reaction processing and optical measurement consistently with one device using one or more types of magnetic particles. An object of the present invention is to provide a magnetic particle reaction control device using a variable pitch dispensing device that is highly efficient and can perform processing quickly, and a reaction control method thereof.

The third object is to provide a device that can process magnetic particles when containing wells or the like arranged at various pitches without complicating the device structure and increasing the manufacturing cost. Accordingly, it is an object of the present invention to provide a magnetic particle reaction control device and a reaction control method using a variable pitch dispensing device that can be executed without complicating the processing.

According to a first aspect of the present invention, there are provided a plurality of nozzles arranged at a variable pitch in at least one row having tip portions capable of sucking and discharging a liquid, and at least the first pitch and the second in accordance with instructions from the nozzles. A nozzle head having a pitch conversion mechanism capable of converting into a pitch and a plurality of nozzle heads that can be inserted all at once and arranged in one or more rows along the row direction at the first pitch. A first housing portion group having housing portions and a second housing having a plurality of housing portions arranged in one or more rows along the row direction at a second pitch smaller than the first pitch. A group of units, a moving mechanism that allows the nozzle head or nozzle to move relative to the first storage unit group and the second storage unit group, and at least the first pitch or the second pitch Arranged A magnetic device capable of simultaneously applying and removing a magnetic field in the tip portion of the nozzle, and along the column direction in either the first housing unit group or the second housing unit group It is a magnetic particle reaction control device using a variable pitch dispensing device in which a magnetic particle suspension is accommodated in an arrayed accommodating portion.

Here, the “variable pitch” means that when the same shape (for example, a container, a well, a nozzle, etc.) is arranged at equal intervals, the interval of the arranged ones is variable by an instruction. .

The “nozzle” is a portion where fluid is sucked and discharged, and the fluid includes gas and liquid. The nozzle is a flow path that communicates with a cylinder having a plunger, a pump, or a mechanism that sucks and discharges gas by deformation of a bellows or an elastic body. The nozzle also includes a dispensing tip attached to a mounting nozzle (a fitting portion thereof) as a tip portion. The “plurality of nozzles” is preferably equal to or equal to the number of accommodating portions arranged in the row direction of the first accommodating portion group and the second accommodating portion group, for example.

The “first pitch” and “second pitch” depend on the amount of liquid to be handled, the size and shape of the container that accommodates it, the pitch of the arrangement of the containers, the pitch of the wells of the standardized microplate, etc. Determined. There may be a housing group having a third pitch or the like other than this. The “accommodating group” may contain an array of containers, the microplate or cartridge container group. When the nozzles of the nozzle head are set to a predetermined variable pitch, the phase or reference position of the array is provided with a fixed nozzle (to be described later) that is fixed by pitch conversion. The fixed nozzle is used as a reference for the array, and the fixed nozzle is inserted. Each accommodating part such as possible corresponding well, liquid accommodating part, chip accommodating part and the like is provided along a straight movement path of the stationary nozzle, and arranged in the row direction at the same pitch as that pitch. Is preferred. Thereby, the movement can be smoothly performed in a straight line along the installation direction of the accommodating portion group. There may be a plurality of types of arrangement phases or reference positions depending on the pitch. For example, this is a case where the accommodating unit groups themselves are arranged in a matrix (two directions, the row direction and the column direction).

“Magnetic particles” are particles having magnetism and have a size of, for example, about 1 nm to several tens of μm. The size, mass, material, structure (single domain, surface coated with various coating substances, etc.), properties (paramagnetism, superparamagnetism, ferromagnetism, ferrimagnetism, magnitude of magnetic force), etc. are processed. It can be determined according to the purpose. The material is composed of iron hydroxide, iron oxide hydrate, iron oxide (γ-Fe ₂ O ₃ , Fe ₃ O ₄ etc.), mixed iron oxide, or iron. Magnetic particles can be obtained by coating the material with various coating materials. Coating materials include organic substances that generate various functional groups, ionic substances that generate ions, and surface stabilizing substances that prevent aggregation and precipitation due to magnetic fields (aliphatic diols, polycarboxylic acids and their substitution products and derivatives). Etc.), specific binding substances (ligands, receptors, etc.), medicinal active substances, etc. Alternatively, magnetic particles can be magnetized by attaching, incorporating, or bonding a magnetic substance to a non-magnetic carrier, for example, an inorganic substance such as silica, glass, ceramics, metal, or an organic substance such as cellulose, agarose gel, rubber, or nylon. You may make it use as.

A “ligand” is a molecule that is bound by a specific receptor, and includes, for example, genetic materials such as nucleic acids, and biological materials such as proteins, sugars, sugar chains, and peptides. For example, agonists and antagonists for cell membrane receptors, toxins (toxin and venom), viral epitopes, hormones, hormone receptors, peptides, enzymes, enzyme substrates, lectins, sugars, oligonucleotides, polynucleotides, oligosaccharides, antibodies, etc. . Natural or artificial materials may be used. The “receptor” has binding ability to the ligand, and includes, for example, genetic materials such as nucleic acids, and biological materials such as proteins, sugars, sugar chains, and peptides. More specifically, examples of a combination of a ligand and a receptor include various antigens and antibodies, for example, biotin and avidin, biotin and streptavidin, and the like.

The first container part group or the second container part group is, for example, a cartridge container in which a plurality of cartridge containers equal to or more than the number of the nozzles are arranged along the row direction at the first pitch or the second pitch. It is a microplate in which a plurality of wells equal to or more than the number of the nozzles are arranged in groups along the row direction.

“Microplate” refers to a container in which a predetermined number of wells (liquid storage portions) are arranged in a matrix at a predetermined pitch (row pitch and column pitch). Among them, standardized microplates include, for example, 12 rows x 8 columns 96 well microplates, 24 rows x 16 columns 384 well microplates, 48 rows x 32 columns 1536 well microplates. The pitches are 9 mm, 4.5 mm, and 2.25 mm, respectively. Examples of the material for the microplate include resins such as polyethylene, polypropylene, polyester, polystyrene, polyvinyl, and acrylic.

“Cartridge container” means a liquid container that can store liquid, a chip container that can store dispensing tips, a tube-shaped liquid container or a hole that can hold a chip container, a temperature-controllable reaction container, etc. The accommodating portions are arranged in a line. When a plurality of cartridge containers are arranged at a predetermined pitch in the column direction, each cartridge container extends in the row direction.

The “row direction” refers to the arrangement direction of the plurality of nozzles as a row direction (corresponding to the X-axis direction in the embodiment) for convenience. Among the “multiple columns”, elements, for example, wells or nozzles, are arranged in a predetermined row pitch and a column pitch along a column direction and a row direction, respectively. The structure is called “matrix”, and the number of rows and the number of columns is 2 or more, respectively. The column direction and the row direction are usually orthogonal, but are not necessarily limited thereto and may be oblique. In the case of a matrix arrangement, the pitch along the column direction is “row pitch”, the pitch along the row direction is “column pitch”, and “arrange in a matrix at the first pitch”. Means that the row pitch along the column direction is equal to the first pitch. Note that the row pitch and the column pitch are not necessarily the same. Since it has “at least”, it is possible to have one or two or more accommodating portions having a third pitch or the like other than “first pitch” and “second pitch”. .

According to a second aspect of the present invention, the magnetic device is provided in the nozzle head, and one or one set of magnets provided corresponding to the nozzles is at least at the first pitch or the second pitch. Corresponding to the magnet rows arranged in at least one row along the row direction and the tip of each nozzle when the variable pitch of the nozzles is set to the first pitch or the second pitch A magnetic particle reaction control device using a variable pitch dispensing device having a magnet row moving mechanism that moves the magnet row so that the magnets can be brought into and out of contact at the same time.

”Here,“ contact and separation ”means accessible and separable. “One or one set of magnets provided corresponding to each nozzle” means one or one set of magnets that exert the most magnetic influence on the nozzles by simultaneous access to the arranged nozzles ( In the case of a set of magnets, a plurality of magnets (these magnets do not necessarily have to have the same shape and magnetic force) are included. In order for one or a set of magnets to exert a homogeneous and strong magnetic force on the tip of each nozzle, the same structure (for example, the included magnets have the same shape, the same or a symmetric shape) Those having an arrangement (positional relationship) or having the same magnitude of magnetic force are preferable. In particular, with respect to the magnetic poles (N pole, S pole) of one or a set of magnets, the repulsive force or attractive force exerted between adjacent magnets of the arranged magnets is relaxed, and the installation of the magnets is stabilized. Preferably, the magnetic poles are alternately reversed between adjacent magnets, or the arrangement of the magnetic poles is alternately reversed between a pair of adjacent magnets (that is, the N poles and the S poles are alternately arranged) ). Note that the total number or the total number of magnets is equal to the number of nozzles when one magnet or one set of magnets applies a magnetic force mainly to one nozzle by the approach. If one magnet or one set of magnets gives the same magnetic force to two or more nozzles, the total number of magnets or the total number of sets may be smaller than the number of nozzles. Thus, when extra magnets are arranged outside the nozzle row in order to maintain the magnetic force uniformity, the total number of magnets or the total number of sets may be larger than the number of nozzles.

In the “magnet row”, for example, magnets are arranged at a predetermined pitch along a straight line (or a curve) that does not intersect with the nozzles (for example, the center of gravity of each magnet is parallel to the row direction of the nozzles). For example, each magnet is fixed to one or a plurality of magnet support members extending in the row direction of one or a plurality of rows of nozzles so as not to move. When a plurality of magnet support members are used, they are attached in a comb-teeth shape to a connection support body connected to the magnet row moving mechanism at one end thereof. The magnet row moving mechanism moves these magnets relatively simultaneously along the row direction or in a direction perpendicular or oblique to the row direction with respect to the nozzles so as to contact and separate.

The “magnet column” is, for example, only movement of the magnet column itself in the row direction without moving the magnet column itself in the column direction (nozzle arrangement direction) even when the variable pitch of the nozzle head is converted. It is desirable that a magnetic field can be applied to the tip of the nozzle.

According to a third aspect of the present invention, in the magnet row, the one or one set of magnets corresponding to the nozzles are arranged in at least one row along the row direction at the first pitch and the second pitch. The magnet row moving mechanism simultaneously contacts and separates the magnets corresponding to the tip of each nozzle when the variable pitch of the nozzles is set to the first pitch and the second pitch. It is a magnetic particle reaction control device using a variable pitch dispensing device that moves the magnet row so as to enable it.

Here, the arrangement of the magnets of the first pitch p1 and the second pitch p2 is one row as at least a common magnet row along the row direction with a magnet corresponding to the nozzle as a common reference with respect to a predetermined nozzle. It is preferable to arrange them together in a shape. In this case, when there is a magnet position common to the first pitch and the second pitch, one of the corresponding magnets can be omitted, so that the first pitch and the second pitch are separated. Compared with the case where the magnet rows are provided, the number of magnets can be reduced and the arrangement of one magnet is sufficient, so that the apparatus scale can be prevented from becoming complicated.

For example, one or a set of magnets may be arranged in the same direction as the same magnet row along the row direction with a common magnet corresponding to a nozzle (for example, a non-moving nozzle to be described later) whose position is not changed by the pitch conversion. When the magnets are arranged in a line with the pitch p1 and the second pitch p2, in order to reduce the number of magnets as much as possible, for example, the first pitch p1 is set to the second pitch p2. The rational number ν times is preferable. In this case, a predetermined common maximum pitch p, which will be described later, is obtained, and the coordinate position corresponding to the first pitch p1 and the second pitch p2 among the coordinate positions in units of p, the number of the nozzles. The magnets corresponding to the number of are arranged. That is, p1 / p2 = a · p / b · p = ν> 1 (a and b are natural numbers such that a> b and are relatively prime. P is called a common maximum pitch). When the first magnet position is common to the first pitch p1 array and the second pitch p2 array, the magnet is common at the coordinate position of the common multiple of a and b. The number can be reduced. In addition, it is preferable to reduce the number of magnets when the rational number (m × ν) of the number of nozzles (m> 1, natural number) is a natural number. Thus, regardless of whether the nozzles are set to the first pitch p1 or the second pitch p2, a magnetic field can be applied to the tip of each nozzle, and the number of magnets can be minimized. become. When the pitch p is large and the influence of the adjacent magnetic force is small, as described above, magnets are provided only at the coordinate positions corresponding to the first pitch p1 and the second pitch p2 among the coordinate positions. Further, it is not necessary to continuously arrange at the respective coordinate positions spaced by the pitch p. On the other hand, when the pitch p is small and the influence of the adjacent magnetic force is large, considering the homogeneity of the magnetic field exerted on each nozzle, for example, over the entire length (m−1) × p1 of the magnet row, It may be preferable to provide magnets continuously with respect to the coordinate positions or at coordinate positions other than the first pitch p1 and the second pitch p2 (one at a time or a suitable one for the sake of homogeneity outside). There may be a case where a number of magnets are provided on each outer side at the pitch p). However, since only one magnet row is required, the number of magnet support members that support the magnet row can be reduced, and the structure of the nozzle head can be simplified. Even in the magnet row in this case, it is preferable to alternately reverse the magnetic poles of the adjacent magnets or the arrangement of the magnetic poles.

In particular, when ν is a natural number n, (n−2) types (n> 2) of pitches other than the first pitch p1 and the second pitch p2 can be set. In this case, p2 is the maximum common pitch p. The number of magnets is, for example, (m−1) × n + 1. When the magnets at both ends of the magnet row correspond to both ends of the tip row of the nozzle, it is preferable to provide at least one magnet further outside the magnets at both ends in order to maintain the homogeneity of the magnetic field. . Accordingly, the number in this case is, for example, (m−1) × n + 3. The arrangement of one or a set of magnets contributes to the simplification of the structure by arranging them with reference to the magnets corresponding to the stationary nozzle described later. In the above description, only the first pitch and the second pitch have been described. However, even in the case where there are more pitches than the first pitch, they are merged into a single row based on the above-described purpose, for example, p3 = c · p (A, b, and c are relatively prime) and can be expanded by obtaining the common maximum pitch p.

According to a fourth aspect of the present invention, the magnetic device is provided in the nozzle head, and at least one row of magnets or a set of magnets provided corresponding to the nozzles is arranged at a variable pitch that is the same as the variable pitch of the nozzles. A variable pitch magnet array arranged in a shape, and a magnet array moving mechanism for moving the magnet array so that the arrayed magnets can be brought into and out of contact with the tip portions of the corresponding nozzles all at once. The variable pitch magnet array is a magnetic particle reaction control device using a variable pitch dispensing device that is pitch-converted by the pitch conversion mechanism together with the nozzle. Even in the magnet row in this case, it is preferable to alternately reverse the magnetic poles of the adjacent magnets or the arrangement of the magnetic poles.

As the variable pitch magnet row, for example, magnets are arranged along a straight line (or curve) that does not intersect with each nozzle (for example, parallel to the arrangement direction of the nozzles) at a predetermined variable pitch (for example, It is provided on the magnet support members arranged at a variable pitch of the number corresponding to the number of the nozzles arranged along the row direction of one or a plurality of rows of nozzles. For example, the variable pitch magnet array is provided with a magnet array moving mechanism which is provided so as to be relatively movable along the row direction orthogonal to the arrangement direction and moved toward and away from the nozzle. In this case, each magnet and each nozzle have a one-to-one correspondence, and the total number or total number N of magnets and the number m of nozzles are equal.

The fifth invention further includes an independent nozzle moving mechanism capable of moving the plurality of nozzles up and down independently from each other with respect to the nozzle head, and the pitch converting mechanism converts the variable pitch of the nozzles to a third pitch. And having a third housing portion group having a plurality of housing portions arranged in one or more rows along the row direction at the third pitch, the third housing The part group is a magnetic particle reaction control device using a variable pitch dispensing device in which sample storage portions capable of storing the collected sample solution are arranged in at least one row along the row direction.

Here, the “third pitch” is not necessarily limited to the case where it is different from the first pitch and the second pitch.
The independent nozzle moving mechanism (Z-axis independent moving mechanism) in which the nozzles can be moved up and down independently of each other is provided because the amount of the sample solution collected in the sample container varies depending on the sample. Therefore, the liquid level of the sample solution stored in each sample storage unit is different. Then, in order to aspirate a common predetermined amount of liquid necessary for the test from the inside of the specimen storage unit, the liquid level of the specimen solution is detected for each nozzle, and after the liquid level is detected, the liquid level is detected for a predetermined time. This is because suction with pressure is required. Therefore, the nozzle is provided with a liquid level detection sensor.

In a sixth aspect of the present invention, the third pitch is larger than the first pitch, and the first storage section group includes a storage section that stores at least one type of reagent solution for reacting the sample solution. Arranged in at least one row along the row direction, and containing the magnetic particle suspensions in at least one row along the row direction, temperature-controllable reaction vessels along the row direction Magnetic particle reaction control using a variable pitch dispensing device in which light measurement wells are arranged in at least one row along the row direction. Device.

As a reaction vessel capable of controlling the temperature, for example, when the variable pitch of the nozzle is set to the first pitch or the second pitch, each of the tip portions of the nozzle can be inserted. And a temperature control block provided with a plurality of insertion portions into which the accommodation portions or wells can be inserted.

According to a seventh aspect of the present invention, each nozzle includes a mounting nozzle and a tip portion that is detachably mounted on the mounting nozzle, and the variable pitch of the nozzle is the first pitch, the second pitch, or When the third pitch is set, the first pitch, the second pitch, or the third pitch in a state where the tip can be mounted on the mounting nozzle by lowering the mounting nozzle. In the magnetic particle reaction control device using the variable pitch dispensing device, the tip portion accommodating portion that accommodates the tip portion further includes a tip portion accommodating portion group arranged in at least one row along the row direction.
In addition, it is preferable to further provide a detachable member for detaching the tip portion on the nozzle head or the stage. Here, the “tip portion” corresponds to a dispensing tip to be described later.

In an eighth aspect of the invention, the nozzle includes a stationary nozzle that is stationary when the variable pitch of the nozzle by the pitch variable mechanism is converted into at least a first pitch and a second pitch, and the first housing The arrangement of the part group and the second accommodation part group is such that the stationary nozzle corresponding accommodation part into which the tip of the stationary nozzle is to be inserted is arranged along the linear movement path of the stationary nozzle by the moving mechanism. Each of the magnets in the magnet array is a magnetic particle reaction control device using a variable pitch dispensing device arranged with reference to a position corresponding to the stationary nozzle.

The number of the nozzles along the column direction when the stationary nozzle corresponding accommodating portions into which the stationary nozzles are to be inserted are arranged along the linear movement path of the stationary nozzles (corresponding to the “row direction”). When the plurality of accommodating portions equal to each other are arranged, the tip of the nozzle can be inserted into each accommodating portion only by relative movement of the nozzle head in the row direction without moving the nozzle head in the column direction. It becomes.

According to a ninth aspect of the present invention, there is provided a variable pitch of the nozzles of a nozzle head in which a plurality of nozzles having tip portions capable of sucking and discharging liquid are arranged in one or a plurality of rows at a variable pitch that can be converted by an instruction. A first accommodation section group having a plurality of accommodation sections arranged in one or a plurality of rows along the row direction at the first pitch, and a first pitch setting step of setting to the first pitch A first suction / discharge step of performing suction or discharge by moving the nozzle head or nozzle relatively, and moving the nozzle head or nozzle relative to the first housing portion group. A second pitch setting step of extracting the tip of the nozzle from the housing and setting the variable pitch of the nozzle to a second pitch smaller than the first pitch; Or multiple rows A second suction / discharge step of performing suction or discharge by moving the nozzle head or nozzle relative to a second storage portion group having a plurality of storage portions arranged, and Magnetic particle suspensions are accommodated in the accommodating portions arranged in at least one row along the row direction in either the first accommodating portion group or the second accommodating portion group, and the first suction / discharge step or By applying a magnetic field in the tip of each nozzle arranged in at least one row in the row direction at the first pitch or the second pitch at the time of suction or discharge in the second suction / discharge step, A magnetic field separation step for adsorbing and separating the magnetic particles on the inner wall of the tip, and removing the magnetic field from the tip of each nozzle during the suction or discharge, thereby causing the magnetic particles to be adsorbed on the inner wall of the tip. This is a magnetic particle reaction control method using a variable pitch dispensing device that further includes a resuspension step in which the magnetic particles are detached from the inner wall and resuspended in the liquid.

In a tenth aspect of the invention, the magnetic field separation step includes at least one magnet or a set of magnets corresponding to each nozzle in the suction or discharge in the first suction / discharge step or the second suction / discharge step. The magnet row arranged in the row direction in at least one row at the first pitch or the second pitch is moved to move the one or one set of magnets to the tip portion of each corresponding nozzle. A magnetic field is applied to the inside of each tip by approaching all at once, and the resuspension step moves the magnet row during the suction or discharge through the tip of the nozzle, This is a magnetic particle reaction control method using a variable pitch dispensing device which is performed by separating the corresponding one or one set of magnets from the tip of each nozzle at once.

In an eleventh aspect of the present invention, at the time of the suction or discharge in the first suction / discharge step and the second suction / discharge step, at least the first pitch and one set of magnets corresponding to each nozzle By moving a magnet row arranged in at least one row at a second pitch and bringing the one or one set of magnets close to the tip portions of the corresponding nozzles at the same time, a magnetic field is generated in each tip portion. This is a magnetic particle reaction control method using a variable pitch dispensing device that affects the magnetic field.

In a twelfth aspect of the invention, the magnetic field separation step is provided in the nozzle head during the suction or discharge in the first suction / discharge step or the second suction / discharge step, and corresponds to each nozzle. Each of the one or one set of magnets corresponding to the one or one set of magnets is moved by moving a variable pitch magnet row arranged in at least one row at the same variable pitch as the variable pitch of the nozzle. A magnetic field is applied to the inside of each tip by simultaneously approaching the tip of the nozzle, and the resuspension step is performed during the suction or discharge through the tip of the nozzle. This is a magnetic particle reaction control method using a variable pitch dispensing device that moves the one or a set of magnets away from the tip of each nozzle at the same time.

In a thirteenth aspect of the present invention, prior to the first pitch setting step, a third pitch setting step of setting the variable pitch of the nozzle head to a third pitch larger than the first pitch; The nozzle head or the nozzle is moved relative to a third housing portion group having a plurality of housing portions arranged in one or more rows in the row direction at a third pitch, and each nozzle is moved. And a third aspirating and discharging step for aspirating the collected sample solution accommodated in the accommodating portions arranged in the row direction of the third accommodating portion group while moving up and down independently of each other. In the first suction and discharge step, the first storage section group accommodated in at least one row along the row direction in which at least one kind of reagent solution for reacting the sample solution is accommodated. And the magnetic particle suspension is contained A step of performing suction and discharge with respect to the housing portions of the first housing portion group arranged in at least one row along the row direction, and in the row direction after the second suction and discharge step. A magnetic particle reaction control method using a variable pitch dispensing device having a light measurement step of measuring light with respect to the light measurement wells of the second accommodation unit group arranged in at least one row along is there.

In a fourteenth aspect of the invention, the nozzle includes an attachment nozzle and a tip portion detachably attached to the attachment nozzle, and the nozzle head includes the nozzle head before or in the first pitch setting step. The variable pitch of the mounting nozzle is set to a first pitch, a second pitch, or a third pitch larger than the first pitch, and the tip portion can be mounted on the nozzle head or the mounting nozzle. In this state, the mounting is performed by moving to the tip portion accommodating portion group accommodated in the plurality of accommodating portions arranged at the first pitch, the second pitch, or the third pitch, and lowering the attachment nozzle. It is the magnetic particle reaction control method using the variable pitch dispensing apparatus which has the process of attaching the said front-end | tip part to the nozzle for use.
In addition, it is preferable to have the removal | desorption process which remove | desorbs the front-end | tip part of this nozzle after the installation process which mounts | wears with the front-end | tip part of a nozzle.

In a fifteenth aspect of the present invention, in the first pitch setting step and the second pitch setting step, the pitch among the plurality of nozzles arranged in at least one row along the row direction provided in the nozzle head. With respect to the stationary nozzles that are stationary during the conversion, the first storage unit group and the second storage unit are arranged so that the movement path in the first suction / discharge step and the second suction / discharge step is linear. The stationary nozzle corresponding accommodating portions into which the stationary nozzles in the accommodating portion group are to be inserted are arranged along the movement path, and the magnet arrangement in the separation and extraction step is based on the stationary nozzles. This is a magnetic particle reaction control method using an arrayed variable pitch dispensing device.

According to the first invention or the ninth invention, the first accommodating portion group arranged at least at the first pitch, and the second accommodating portion arranged at the second pitch smaller than the first pitch. In addition, the processing is performed using a variable pitch nozzle head provided with a magnetic device capable of exerting a magnetic force on the nozzles arranged at least at the first pitch or the second pitch. Therefore, with one common nozzle head, it is possible to perform processing while transferring only the target substance using magnetic particles to the storage unit groups having different pitches and leaving unnecessary liquid in each container. Efficient and reliable processing can be performed. In addition, since the nozzle head or the nozzle can be moved to perform processing on the accommodating portion group having the first pitch and the second pitch, the moving mechanism can be simplified to reduce the number of parts and the scale of the apparatus. It can be reduced, manufacturing costs can be reduced, and processing can be performed efficiently and quickly.

Also, the storage units can be arranged and used at a pitch that is easy for the user to handle according to the processing content. For example, for collecting a sample from a patient, a relatively large container that can accommodate an amount in consideration of using the sample for various tests can be used. In order to aspirate a small amount of sample necessary for testing from these containers and perform processing using a small amount of reagents, a large number of containers are arranged and integrated at a relatively small pitch to reduce the work area and improve efficiency. Can be handled.

In this way, by providing containers of various sizes necessary for processing on one stage and using one common nozzle head provided with a magnetic device, reaction control of magnetic particles is consistently and efficiently performed. It can be done quickly.

According to the second invention or the tenth invention, at least one or one set of magnets provided corresponding to each nozzle arranged in one or more rows at the first pitch or the second pitch is provided. A magnet that moves the magnet rows arranged in a row along the row direction at the first pitch or the second pitch so that the magnets can be brought into contact with and separated from the corresponding tip portions of the nozzles all at once. A column moving mechanism is provided in the nozzle head. As a result, the magnets are arranged at intervals of the first pitch or the second pitch without giving a variable pitch to the magnet row and the magnet row moving mechanism even though each nozzle has a variable pitch. By using the magnet array, a uniform and strong magnetic force can be exerted on the nozzles arranged at the first pitch or the second pitch, and the structure of the nozzle head can be simplified.

According to the third invention or the eleventh invention, the one or one set of magnets corresponding to each nozzle is arranged in at least one row along the row direction at the first pitch and the second pitch. Therefore, a uniform and strong magnetic force is exerted on the tip of the nozzle in both cases where the variable pitch of the nozzle is set to the first pitch and the second pitch. be able to. Further, the number of magnets or the number of sets can share the magnets at a common arrangement position, and the number of magnets can be reduced to simplify the structure of the nozzle head.

The first pitch is a natural number multiple of the second pitch (n> 1), and the number of magnets or the number of pairs N is provided as the natural number multiple of the number of nozzles (m). Not only the case of setting to the first pitch and the second pitch but also n types (n> 2) of pitches can be handled.

According to the fourth invention or the twelfth invention, the magnet array moves the variable pitch magnet array in which one or a set of magnets corresponding to each nozzle is arranged in one or more arrays at the same variable pitch as the nozzle. And a magnet row moving mechanism that enables the magnets to be brought into and out of contact with the corresponding tip portions of the nozzles at the same time. Therefore, the number of magnets or the number of sets required is not limited to the number corresponding to the number of nozzles, and the nozzles and magnets need to have a variable pitch, but the magnet array moving mechanism needs to have a variable pitch. Therefore, the structure of the nozzle head is simplified.

According to the fifth invention, by providing a plurality of nozzles that can be moved up and down independently of each other, the liquid volume of the sample solution or the like is indefinite, and the case where the pitch of the container is large, A fixed amount of aspiration can be performed.

According to the sixth invention or the thirteenth invention, the third container having the largest pitch contains the sample solution, and the first container having the next largest pitch contains the reagent used for the reaction of the sample. In addition, a suspension of magnetic particles and a reaction vessel are arranged, and the second container with the smallest pitch is used for measurement. In addition, since the nozzle can be moved up and down independently, it is possible to secure the amount of specimen required for a plurality of examinations and to use for examination by nozzles that move up and down independently at a variable pitch. A simple container can be aspirated and reacted with a reagent, etc., and the process can be performed quickly and smoothly without changing the nozzles consistently until measurement, and an efficient container With this arrangement, processing can be executed without increasing the work space or the scale of the apparatus.

According to the seventh invention or the fourteenth invention, the tip portion of the nozzle is detachably provided, and the tip is provided at the first pitch, the second pitch, or the third pitch so that the tip portion can be attached. It is housed in a part housing part group. Therefore, by moving the nozzle head or the nozzle, the tip of the nozzle can be automatically attached to the nozzle, so that cross contamination is reliably prevented and a plurality of tests are performed on one specimen. Alternatively, a plurality of specimens can be processed in one examination. Moreover, when it accommodates and mounts | wears with an accommodating part with a 1st pitch or a 2nd pitch, the work area on a stage will be reduced more and work efficiency will be improved.

Also, since the tip can be automatically mounted without human intervention, the processing can be performed automatically and consistently.

According to the eighth aspect or the fifteenth aspect, the stationary nozzle is included in one of the nozzles that is stationary when the pitch between the nozzles is changed, so that the movement path of the stationary nozzle is linear. By arranging the nozzle-corresponding accommodating portions along the movement path, the processing is smooth and quick like a nozzle with a fixed pitch regardless of the existence of a plurality of types of accommodating portion groups having different pitches. In addition, the spatial arrangement of the housing parts can be made more efficient, and the structure of the magnetic device, especially the movement mechanism and movement control can be simplified and facilitated, simplifying the device configuration and reducing the device scale. In addition, the work area can be reduced along with the reduction of manufacturing cost.

It is a conceptual perspective view which shows the external appearance of the variable pitch dispensing apparatus which concerns on the 1st Embodiment of this invention. It is a perspective view of the principal part of the variable pitch dispensing apparatus which concerns on the 1st Embodiment of this invention. It is a perspective view of the principal part of the nozzle head which concerns on the 1st Embodiment of this invention. It is a perspective view of the principal part of the magnetic device which concerns on the 1st Embodiment of this invention. It is a perspective view which shows the pitch conversion mechanism in the 1st pitch state which concerns on the 1st Embodiment of this invention. It is a perspective view which shows the pitch conversion mechanism in the 2nd pitch state which concerns on the 1st Embodiment of this invention. It is a perspective view which shows the pitch conversion mechanism in the 3rd pitch state which concerns on the 1st Embodiment of this invention. It is a top view which shows the whole variable pitch dispensing apparatus which concerns on the 1st Embodiment of this invention. It is a perspective view which shows operation | movement of the variable pitch dispensing apparatus which concerns on the 1st Embodiment of this invention. It is a schematic diagram which shows the 1st pitch state of the principal part of the nozzle head which concerns on the 1st Embodiment of this invention. It is a schematic diagram in the case of applying a magnetic field to the dispensing tip in the first pitch state of the main part of the nozzle head according to the first embodiment of the present invention. It is a schematic diagram in the case of applying a magnetic field to the dispensing tip in the second pitch state of the main part of the nozzle head according to the first embodiment of the present invention. It is a perspective view at the time of applying a magnetic field to the dispensing tip of the 1st pitch state of the principal part of the nozzle head concerning a 2nd embodiment of the present invention. It is a side view of the principal part of the nozzle head which concerns on the 2nd Embodiment of this invention. It is a perspective view at the time of applying a magnetic field to the dispensing tip of the 2nd pitch state of the principal part of the nozzle head concerning a 2nd embodiment of the present invention. It is a perspective view at the time of applying a magnetic field to the dispensing tip of the 3rd pitch state of the principal part of the nozzle head concerning a 2nd embodiment of the present invention.

Subsequently, a magnetic particle reaction control device using the variable pitch dispensing device according to the first embodiment of the present invention and a reaction control method thereof will be described with reference to the drawings.

FIG. 1 shows an entire magnetic particle reaction control apparatus 10 using a variable pitch dispensing apparatus according to a first embodiment of the present invention.
The magnetic particle reaction control device 10 using the variable pitch dispensing device has 12 nozzles (corresponding to mounting nozzles) to which a dispensing tip 18 as a tip portion can be mounted, and the pitch between the adjacent nozzles. A pitch conversion mechanism that can be changed by instructions, a magnetic device 20 that can apply a magnetic field to the inside of the dispensing tip 18 mounted on the nozzle, and the nozzle can be moved independently along the Z-axis direction. Nozzle head 70 provided with a Z-axis independent movement mechanism corresponding to an independent nozzle raising / lowering mechanism, a third accommodating portion group 30 having accommodating portions arranged with a third pitch p3, and a third pitch p3 First accommodating portion group 40 having accommodating portions arranged with a smaller first pitch p1 and second accommodating portion group arranged with a second pitch p2 smaller than the first pitch p1. 50 and The twelve nozzles communicate with each other through a flow path, and the suction and discharge of gas to and from each nozzle enables liquid to flow into and out of the dispensing tip 18 attached to the nozzle. Each of the pumps 15 and a real-time PCR method processing apparatus 90 that performs temperature control and optical measurement regarding the processing based on the real-time PCR method performed in the second housing unit group 50 are provided on a stage 99. As a whole, it is incorporated in the housing 98.

A Y-axis moving mechanism that allows the nozzle head 70 to move in the Y-axis direction (row direction) on the stage 91 in the housing 98 of the magnetic particle reaction control device 10 using the variable pitch dispensing device. And a ball screw, a nut portion screwed into the ball screw, a motor for rotating the ball screw, and the like. Each of the accommodating portions of the accommodating portion group 30, 40, 50 includes the nozzles 12 ₁ to 12 so that the dispensing tip 18 attached to the nozzle can pass above the accommodating portions by the Y-axis moving mechanism. 12 ₁₂ are arranged along a movement path in the Y-axis direction. The Y axis moving mechanism corresponds to the moving mechanism of the nozzle head 70, and the combination of the Z axis independent moving mechanism 80 and the Y axis moving mechanism corresponds to the moving mechanism of each nozzle.

FIG. 2 is an enlarged view showing the main part of the magnetic particle reaction control device 10 using the variable pitch dispensing device according to the first embodiment shown in FIG. 1 in more detail.
As described above, the nozzle head 70 includes twelve nozzles 12 ₁ to 12 ₁₂ (corresponding to mounting nozzles), the magnetic device 20, and at least the first pitch p1, the second pitch p2, And a pitch conversion mechanism 60 that enables pitch conversion between the third pitches p3. The nozzles 12 ₁ to 12 ₁₂ are provided with a flow path 13 above them, and the switching valves 16 of the pumps 15 are provided. Gas communicates with the cylinder 14 through the interior. A plunger is slidably provided in the cylinder 14, and a reference numeral 17 is a pump drive unit in which a circuit board of the pump 15 is built. A fitting portion 12a is provided at the lower end of each of the nozzles 12 ₁ to 12 ₁₂ and can be connected by fitting with the mounting opening portion 18f of each dispensing tip 18. Each of the nozzles 12 ₁ to 12 ₁₂ is provided with a pressure sensor, and can measure the pressure in the dispensing tip 18 attached to detect the liquid level in the container (for explanation, A state in which one dispensing tip 18 is attached is shown, but twelve dispensing tips 18 can be attached all at once).
Here, for reasons described later, the first pitch is set to n times (here, twice) the second pitch, and the third pitch p3 is the first pitch p1 and the second pitch. It is an arbitrary pitch unrelated to the pitch p2.

Each of the nozzles 12 ₁ to 12 ₁₂ is held in a vertical hole that vertically penetrates the support plates 81 ₁ to 81 ₁₂ in the vertical direction (Z-axis direction), and the 12 support plates 81 ₁ to 81 ₁₂ are arranged in the thickness direction. Are arranged so as to extend along the X-axis direction, and guide rails 66a penetrating through the support lateral holes respectively drilled along the X-axis direction so as to penetrate the thicknesses of the _twelve support plates 81 ₁ to 81 ₁₂ is slidably supported by 66b, the rails 66a for the guiding one end of the support plate 81 ₁ side 66b is supported by a side plate 73 provided on the substrate 71 of the nozzle head 70, the other end, later as to, and it is supported and supported by the fixed support plate 81 ₁₂ itself on the substrate 71.

As will be described later, the adjacent support plates 81 ₁ to 81 ₁₂ are coupled to each other at a predetermined node provided on the links 61 and 62 via twelve links 61 or 62. Further, among the _twelve support plates 81 ₁ to 81 ₁₂ , eleven support plates 81 ₁ to 81 ₁₂ excluding the support plate 81 ₁₂ are moved in the X-axis direction by a timing belt 65 b and a timing belt 65 b. Two pulleys 65a are provided so as to be supported by the substrate 71 of the nozzle head 70.

The nozzle head 70 is provided with a magnetic device 20 below the _twelve support plates 81 ₁ to 81 ₁₂ . The magnetic device 20 has 23 magnets 22 arranged at a second pitch along the X-axis direction while alternately reversing the magnetic poles (that is, alternately on the side where the S pole and N pole face the nozzle). The magnet rows arranged (in a line), the magnet support block 25 as a magnet support member for fixing and holding the magnet rows, and the dispensing tips 18 attached to the nozzles 12 ₁ to 12 ₁₂ A shaft 26b extending in the Y-axis direction capable of advancing and retreating the magnet rows so that the magnet rows can be simultaneously approached or separated from each other along the Y-axis direction with respect to the narrow tube 18c, and a substrate of the nozzle head 70 A linear motor 27 that is attached to a wall 72 supported by 71 and drives the magnet 22 along the Y-axis direction via the shaft 26b; and 2 that is attached to the wall 72 and extends along the Y-axis direction. Book guide And a rod 26a. A detachable member 21 that protrudes from the magnetic pole surface of the magnet 22 and allows the dispensing tip 18 to be detached from the nozzles 12 ₁ to 12 ₁₂ is provided below the magnet support block 22. . Here, the shaft 26b, the linear motor 27, and the guide rod 26a correspond to the magnet row moving mechanism.

First housing portion group 40 having the first pitch p1 is 12 of the pipette tip 18 in the top fitting opening 18f, the nozzle 12 ₁ by downward movement of the nozzle _{12 1-12 12} 12 to 12 ₁₂ as a tip portion accommodating portion group having twelve hole portions 42 a arranged at the first pitch p 1 in the X-axis direction so that the fitting portion ₁₂ a at the lower end can be fitted and attached. The chip rack 42 includes a group of cartridge containers 41 in which twelve cartridge containers 41 formed so as to extend in the Y-axis direction are arranged along the X-axis direction at a first pitch p1. Each cartridge container 41 includes a various reagent storage unit group 44 having a well 43 in which various reagents are stored or can be stored, and a reaction container group 46 having a reaction container of each capacity. Here, the first pitch p1 is, for example, 18 mm.

The third storage unit group 30 having the third pitch p3 includes a sample storage unit 31 such as 12 blood collection tubes and a third pitch along the X-axis direction of the 12 sample storage units 31. It has a sample storage unit group 32 arranged in p3. Here, the third pitch p3 is, for example, a 22 mm pitch.

The second accommodating portion group 50 having the second pitch p2 has a microplate 51 in which wells 52 of 8 rows × 12 columns are arranged in a matrix. The second pitch is 9 mm, for example. Therefore, in this case, the predetermined common maximum pitch p of the first pitch p1 and the second pitch p2 corresponds to 9 mm, and corresponds to p1 / p2 = 2 (natural number).

FIG. 3 shows a Z-axis independent moving mechanism 80 that can raise and lower the ₁₂ nozzles 12 ₁ to 12 ₁₂ provided in the nozzle head 70 independently along the Z-axis direction. Each of the twelve support plates 81 ₁ to 81 ₁₂ has vertical holes vertically passing through the nozzles 12 ₁ to 12 ₁₂ and held therein, and the support plates are provided on both sides of the vertical holes. Six horizontal holes 82 that pass through 81 ₁ to 81 ₁₂ in the X-axis direction are arranged so as to have translational symmetry so that they do not contact the vicinity along the outside of the vertical holes. Note that only the two support plates 81 ₁ and 81 _{12 at} both ends have a plate shape different from the other ten support plates 81 ₂ to 81 ₁₁ . In a state where these support plates 81 ₁ to 81 ₁₂ are slidably supported on the guide rails 66a and 66b, the same Y-axis coordinates and Z-axis coordinates provided on the support plates 81 ₁ to 81 ₁₂ are specified. The horizontal holes 82 are arranged so as to be coaxial along the X-axis direction. Therefore, the twelve hexagonal shafts 85 allow the _twelve support plates 81 ₁ to 81 ₁₂ to pass through the horizontal holes 82.

On the surface of each of the 12 cylindrical nozzles 12 ₁ to 12 ₁₂ held by the support plates 81 ₁ to 81 ₁₂ , peaks and valleys as teeth of the rack 84 are repeatedly formed in parallel along the circumferential direction. Has been. Also made to correspond to those in the one in a in different for each of the support plate _{81 1-81 12} positions of 12 of the transverse hole 82 provided in the support plate _{81 1-81 12,} in its transverse bore, A coaxial hole having an inner diameter larger than the inner diameter of the horizontal hole and partially overlapping with the vertical hole is formed, and a pinion 83 engaged with the rack 84 is rotatably accommodated therein. A hexagonal cylindrical lateral hole is coaxially formed in the central portion of the pinion 83 and can be fitted to the 12 hexagonal shafts 85. As shown in FIG. 3, twelve hexagonal shafts 85 are provided so as to penetrate twelve lateral holes 82 along the X-axis direction. These hexagonal shafts 85 are connected to twelve different motors (not shown) and are independently rotatable. Accordingly, by rotating any of the hexagonal shafts 85, the nozzles 12 ₁ to 12 ₁₂ provided with the rack 84 held by the support plates 81 ₁ to 81 ₁₂ provided with the corresponding pinions 83 are moved in the vertical direction. Will be moved to.

The dispensing tip 18 has a thick pipe 18b capable of storing the introduced liquid, and the inflow of liquid through the mouth 18a that is formed in a narrower shape than the thick pipe 18b in communication with the thick pipe 18b. And a narrow tube 18c capable of flowing out, and a transition portion 18e connecting the thick tube 18b and the thin tube 18c, and the mounting opening 18f is provided on the upper side of the thick tube 18b, Are provided with a plurality of protrusions 18d extending in the vertical direction so as to project outward from the thick tube 18b. The inner diameter of the hole portion 42a of the tip rack 42 is smaller than the outer diameter of the protrusion 18d or the mounting opening 18f and larger than the outer diameter of the thick tube 18b.

FIG. 4 shows in detail the magnetic force device 20 which is the nozzle head 70 and is provided below the _twelve support plates 81 ₁ to 81 ₁₂ .
The magnetic device 20 has 25 magnet rows 22, 22a, 22b arranged at a second pitch p2 along the X-axis direction (row direction). This is because when the first pitch p1 is n times (in this example, twice) the second pitch p2, m (in this example) for both the first pitch p1 and the second pitch p2. 12) in order to be able to exert a magnetic field homogeneously in the dispensing tips 18, at least the number of magnets need not be (m−1) × n + 1, In order to achieve homogeneity in the magnetic field at both ends of the m dispensing tips 18 (nozzles), at least one magnet is arranged on the outside thereof. In FIG. 3, a magnet 22a and a magnet 22b are magnets for adjusting the magnetic field of the nozzles 12 ₁ and 12 ₁₂ at both ends. Therefore, (m−1) × n + 3 magnets are required for the magnet array. In this case, the length from end to end of the magnet array is {(m−1) × n + 2} × p2. Therefore, when m = 12, n = 2, the total number of magnet rows 22, 22a, 22b is 25, and the length from end to end of the magnet row is 24p2 or 24p. Even in the magnet row in this case, it is preferable to alternately reverse the magnetic poles of the adjacent magnets or the arrangement of the magnetic poles.

As shown in FIG. 4, the magnet rows 22, 22a, 22b are provided in a magnet support block 25 as a magnet support member, and the magnet support block 25 is moved in the Y-axis direction by the drive shaft 26b. The nozzles 12 ₁ to 12 ₁₂ are provided so as to be able to advance and retreat along the nozzles 12 ₁ to 12 ₁₂ and can be brought into contact with and separated from the thin tubes 18c of the dispensing tips 18. A liquid dripping prevention plate (FIG. 6) for preventing liquid dripping from the mouth portion 18a at the tip of the dispensing tip 18 mounted on the nozzles 12 ₁ to 12 ₁₂ is disposed above the magnet support block 25. , 7) may be provided.

As described above, the plate-shaped detachable member 21 is provided on the lower side of the magnet row of the magnetic device 20, and is provided so as to be able to contact and separate from the dispensing tip 18 by the shaft 26b. On the plate of the detachable member 21, semicircular cutouts 23 are arranged in a comb shape at the position of the magnet 22 having the first pitch p1. The inner diameter of the notch 23 is smaller than the outer diameter of the mounting opening 18f of the dispensing tip 18 or the outer diameter of the protrusion 18d formed on the outer surface of the mounting opening 18f. The nozzles 12 ₁ to 12 ₁₂ are formed larger than the outer diameter. Accordingly, the said dispensing tip 18 by moving the cutout portion 23 in advance is advanced to a position to contact the surface of the nozzle _{12 1-12 12,} the nozzles ₁₂ 1 to _{12 12} upwards The nozzles 12 ₁ to 12 ₁₂ can be detached from the nozzles 12 ₁ to 12 ₁₂ .

Next, FIG. 5 shows a pitch conversion mechanism 60 provided in the nozzle head 70.
The pitch conversion mechanism 60, of the guide rail 66a, the support plate supported to 66b ₈₁ 1 ~ _{81 12,} the support plate _{81 12} supports a stationary nozzle _{12 12} unchanged position in pitch conversion, This is the reference position for the entire system including the nozzle head 70. The support plate _{81 12} other supporting plate _{81 1-81 11} except is slidably supported along the X-axis direction. These support plates 81 ₁ to 81 ₁₂ are connected via the twelve links 61 and 62 so that the pitch can be changed as described above.

In the link 61 (length a + 2α) formed shorter than the link 62 (length 2a + 2α), two nodes 63 and 64 are provided at the respective ends at a predetermined distance a, and the node 63 is connected via a connector 69. Te is coupled with the support plate 81 ₁ and the turning pair, are coupled with adjacent links 62 and turning pair at node 64. The link 62 has two nodes 64 of both ends in the middle of the node 63, and from the nodal point 63 to the predetermined distance a, coupled with the support plate 81 ₂ and the turning pair with the node 63, one of the nodes 64 Thus, the link 61 is coupled with a turn pair, and the other node 64 is coupled with the next adjacent link 62 with a turn pair. Similarly, the interval between the adjacent support plates 81 ₁ to 81 ₁₂ is variably connected. Node 64 of the tenth link 62 is attached at around and through the link 61 and the node 64 of the two second kinematic pair, the other nodes 63 in the nodal point 64 at a predetermined distance a in the support plate 81 ₁₂ and turning pairs Are connected.

The support plate 81 ₁ is connected at its lower end to the movable plate 67, the movable plate 67 is connected to the timing belt 65b, it is movable along the X-axis direction by the rotation of the pulley 65a. Reference numeral 65c denotes a motor that rotationally drives the pulley 65a. The support plate 81 _12, the mounted immovably plate 68 which is connected to the substrate 71 of the nozzle head 70, the position of the X-axis direction is fixed to the nozzle head 70. FIG. 5 corresponds to the case where the distance between the nozzles 12 ₁ to 12 ₁₂ is the first pitch p1, here 18 mm pitch.

FIG. 6 shows the support plate and the link when the pitch converting mechanism 60 provided in the nozzle head 70 sets the pitch of the nozzles 12 ₁ to 12 ₁₂ to the second pitch (in this case, 18 mm pitch). The states of 61 and 62 and the movable plate 67 are shown. Note the position of the substrate 71 and the stationary plate 68 has no change, therefore immovable nozzles 12 _12, when the pitch conversion has been shown to be immobile.

FIG. 7 shows the support plate and the link when the pitch converting mechanism 60 provided in the nozzle head 70 sets the pitch of the nozzles 12 ₁ to 12 ₁₂ to the third pitch (22 mm pitch in this case). The states of 61 and 62 and the movable plate 67 are shown. Position without change of the substrate 71 and the stationary plate 68, thus, stationary nozzle 12 ₁₂ has been shown to be still immobile.

FIG. 8 shows an overall plan view of the magnetic particle reaction control device 10 using the variable pitch dispensing device according to the first embodiment of the present invention. Since the nozzle head 70 of the variable pitch dispensing device can move only in the Y-axis direction with respect to the housing group 30, 40, 50, nozzles corresponding to the stationary nozzles arranged in the nozzle head 70 12 ₁₂ X-coordinate position (reference X coordinate position) does not vary by changing the pitch, on the movement path along the Y-axis direction of the nozzle 12 ₁₂ as stationary nozzles (along the row direction), the as immobile nozzle corresponding accommodating portion, the receiving portion of the first pitch of the first housing as the unit group 40-tip end portion housing part group of the chip rack 42 of the nozzle 12 ₁₂ Note min to be attached to the chip 18, the A sample storage section 31 ₁₂ such as a blood collection tube to be sucked by the nozzle 12 ₁₂ of the third storage section group 30 having a pitch of 3, and a first to be sucked and discharged by the nozzle 12 ₁₂ Housing portion having a cartridge chamber _{41 12} of the accommodating portion group 40, housing portion columns _{51 12} of well 52 of the microplate 51 of the second housing part group 50 of the second pitch to be sucked and discharged by the nozzles _{12 12} It is arranged to come. That is, in FIG. 8, the accommodating portions 31 ₁₂ , 41 ₁₂ , and 51 ₁₂ correspond to the stationary nozzle corresponding accommodating portions.

In FIG. 8, as described above, each unused dispensing tip 18 at the first pitch p1 is placed in the first accommodating portion group 40 by the lowering of the nozzles 12 ₁ to 12 _12. A chip rack 42 having holes 42a for accommodating the openings in a row along the X-axis direction with the mounting openings 18f on the upper side so that they can be mounted on 12 ₁ to 12 ₁₂ ; Twelve cartridge containers 41 arranged along the X-axis direction at a pitch p1 are provided, and the cartridge container 41 includes a reagent storage unit group 44 and a reaction container group 46.

The reagent container part group 44 of the cartridge container 41 mainly contains a solution for separation and extraction. The container part 44a contains 40 μl of Lysis 1 and the container part 44b contains 200 μl of Lysis 2. 44c is a binding buffer solution (NaCl, SDS, isopropanol) 500 μl, the storage unit 44d is a first magnetic particle suspension coated with silica capable of capturing nucleic acids (DNA, etc.), and the storage unit 44e , 700 μL of cleaning solution 1 (NaCl, SDS, isopropanol), 700 μL of cleaning solution 2 (50% water, 50% isopropanol) in the storage unit 44 f, and 500 μL of distilled water as a dissociation solution in the storage unit 44 g The portion 44h contains 500 μl of distilled water, and the storage portion 44i stores and stores the second magnetic particle suspension in which a base sequence complementary to the target nucleic acid capable of capturing the target nucleic acid is fixed. The part 44j includes DN A hybridization reagent such as A is accommodated. The reaction container group 46 is temperature-controllable, and has a reaction container holding hole 46a and a reaction container 46b. Under the temperature control, the insertion portions are arranged at a first pitch. A block is provided so that the reaction vessel 46b and the reaction tube can be inserted. In addition, the said isopropyl alcohol (isopropanol) is used for a protein removal as a part of solution for protein separation extraction.

The second accommodating section group 50 includes the microplate 51, a carriage 53 that conveys the microplate 51 to the real-time PCR processing apparatus 90 along the Y-axis direction, and a rail that guides the movement of the carriage 53. 54.
The well row 52a of the microplate 51 contains the cleaning solution 3, and the well row 52b contains a dissociation solution (alkaline solution) for dissociating double-stranded nucleic acids into single strands, and the well row 52c. In the well row 52d, 70 μL of a master mix (SYBR (registered trademark) Green Mix) composed of a real-time amplification reagent, for example, an enzyme, a buffer, a fluorescently labeled primer, and the like is stored. The well columns 52e to 52h are empty well columns, and the real-time PCR processing apparatus 90 can control the temperature.

As described above, in the third storage unit group 30, the sample storage unit group in which the sample storage units 31 such as blood collection tubes are arranged in a line along the X-axis direction at the third pitch p3. 32 is provided. If necessary, it is preferable to have a disposal tank provided so that a disposal port for discarding unnecessary liquid is not affected by the conversion of the variable pitch. For example, the specimen storage unit 31 is a container for storing urine, sewage collected at various places, etc. in addition to whole blood directly collected from 12 subjects.

Although not shown, the present apparatus 10 includes a pump 15 as a suction / discharge mechanism, a pitch conversion mechanism 60, a nozzle head 70 having a Z-axis movement mechanism, the magnetic device 20, a temperature controller, and a Y-axis. A control unit for controlling the moving device and the like is included. The control unit includes, for example, an information processing device including a CPU and a memory, a data input display device such as a mouse, a keyboard, a liquid crystal panel, and a touch panel, a data output device such as a printer, a communication unit, or an external memory such as a CD and a DVD. The drive device etc. are provided.

Next, the operation of the magnetic particle reaction control device 10 using the variable pitch dispensing device according to the first embodiment of the present invention will be described based on FIGS. 8 to 12.
In step S1, the variable pitch of the nozzle head 70 is set to the first pitch p1 (in this example, for example, 18 mm) using the pitch conversion mechanism 60. At this time, the pitch converting mechanism 60 is attached to the timing belt 65b by driving the motor 65c and rotating the pulley 65a to run the timing belt 65b when instructed by the control unit. By moving the movable plate 67, the support plates 81 ₁ to 81 ₁₂ are moved along the X-axis direction, and the variable pitch between the nozzles 12 ₁ to 12 ₁₂ is set to the first pitch p1. At that time, the nozzle 12 ₁₂ is a stationary nozzle does not move by the pitch conversion is fixed immovably plate 68. X-axis coordinate position of the nozzle 12 ₁₂ corresponds to the X-axis reference position.

Next, in step S2, the nozzle head 70 is moved by the Y-axis moving mechanism, and the twelve nozzles 12 ₁ to 12 ₁₂ and their fitting portions 12a move along the Y-axis direction to move the first. It moves to above the chip rack 42 provided in the storage unit group 40.

In step S3, as the Z-axis independent movement mechanism 80, the twelve horizontal holes 82 provided in the twelve support plates 81 ₁ to 81 ₁₂ of the nozzle head 70 are provided so as to pass through. The twelve motors respectively connected to the hexagonal shaft 85 are rotationally driven all at once, so that one different horizontal hole 82 is provided for each of the support plates 81 ₁ to 81 _{12 that} are fitted and connected to the hexagonal shafts 85. The pinion 83 provided in the inside is rotated, meshed with the rack 84 provided in the nozzles 12 ₁ to 12 ₁₂ corresponding to the pinion 83, and the nozzles 12 ₁ to 12 ₁₂ are lowered all at once. by inserting the nozzle 12 _{1-12 12} distal the fitting portion 12a of the fitting opening 18f of the tip rack 42 unused each dispensing chip 18 accommodated in the instrumentation Make.

In step S4, the respective hexagon shafts 85 of the nozzle head 70 are reversely rotated to mesh with the racks 84 provided on the nozzles 12 ₁ to 12 ₁₂ corresponding to the pinions 83 fitted to the hexagon shafts 85, The nozzles 12 ₁ to 12 ₁₂ are raised all at once, and the lower ends of the dispensing tips 18 attached to the nozzles 12 ₁ to 12 ₁₂ are positioned above the tip rack 42.

Next, in step S5, the variable pitch of the nozzle head 70 is converted to a third pitch p3 (in this example, for example, 22 mm) using the pitch conversion mechanism 60. At that time, when there is an instruction from the control unit, the pitch converting mechanism 60 drives the motor 65c to rotate the pulley 65a to run the timing belt 65b and move the timing belt 65b. By moving the plate 67, the support plates 81 ₁ to 81 ₁₂ are moved along the X-axis direction to convert the variable pitch between the nozzles to the third pitch p3.

In step S6, the nozzle head 70 is moved in the Y-axis direction by the Y-axis moving mechanism, and the twelve dispensing tips 18 are positioned above the sample storage units 31 arranged at the third pitch. Move up.

In step S7, the twelve hexagonal shafts 85 provided so as to penetrate through the twelve horizontal holes 82 provided in the twelve support plates 81 ₁ to 81 ₁₂ of the nozzle head 70 are respectively connected. By rotating and driving twelve motors at once, the pinion 83 provided in one different lateral hole 82 is rotated for each of the support plates 81 ₁ to 81 ₁₂ that are fitted and connected to the hexagonal shafts 85. The nozzles 12 ₁ to 12 ₁₂ are engaged with the racks 84 provided in the nozzles 12 ₁ to 12 ₁₂ corresponding to the pinions 83, and the nozzles 12 ₁ to 12 ₁₂ are lowered at the same time in the specimen storage unit 31 such as the blood collection tube The thin tube 18c of the dispensing tip 18 is inserted.

At that time, the pressure sensor provided in each of the nozzles 12 ₁ to 12 ₁₂ detects the pressure of each dispensing tip 18 inserted into the specimen storage unit 31 such as the blood collection tube, thereby the blood collection tube or the like. The capillary 18c of the dispensing tip 18 is lowered independently of the other dispensing tips 18 while detecting the liquid level in the specimen accommodating portion 31. This is because the whole blood collected from the subject is accommodated in each specimen storage unit 31, but the volume thereof varies, and the liquid level is different for each specimen storage part 31. Therefore, in order to use a constant volume of whole blood necessary for processing for each dispensing tip 18, the pump 15 is fixed for a certain period of time after the liquid level is detected for each specimen container 31 such as each blood collection tube. This is because it is necessary to suck a predetermined amount of whole blood by driving with pressure. Then, as shown in FIG. 9A, each dispensing tip 18 is lowered until the liquid level is detected, and thus the height of each dispensing tip 18 is different. In this step, since the magnetic device 20 is not used, the magnet is in a state of being separated from each dispensing tip 18.

The liquid level is detected when, for example, the pump 15 is used to set the pressure in the dispensing tip 18 to a negative pressure, and the mouth 18a at the tip of the dispensing tip 18 comes into contact with the liquid level. This is done by detecting changes in pressure.

In step S8, the pump 15 is driven, and the pump 15 sucks the gas from the nozzle so that a certain amount of whole blood in the pipette tip 18 is sucked. The hexagonal shaft 85 is rotated in accordance with the specimen storage unit 31 such as a blood vessel) to raise the dispensing tip 18 to a certain position above the specimen storage unit 31.

In step S9, using the pitch converting mechanism 60, the interval between the dispensing tips 18 attached to the nozzles 12 arranged at the third pitch p3 and holding the whole blood in the thin tubes 18c and the large tubes 18b. To the first pitch p1. At this time, when instructed by the control unit, the pitch conversion mechanism 60 drives the motor 65c to rotate the pulley 65a to travel the timing belt 65b for a predetermined distance and attach it to the timing belt 65b. The movable plate 67 is moved in the reference X-coordinate position direction to move the support plates 81 ₁ to 81 ₁₂ along the X-axis direction, thereby changing the variable pitch from the third pitch p3 to the first pitch p1. Pitch conversion is performed until

FIG. 10 schematically shows a state in which the variable pitch between the dispensing tips 18 is converted to the first pitch p1. Here, the magnet support block 25 includes 23 magnets 22 arranged at the second pitch p2, and adjusting magnets 22a for adjusting the homogeneity of the magnetic field exerted on each dispensing tip 18. 22b has a magnet row provided on the both sides at the same second pitch as the magnet 22 (in FIG. 10, the dotted rectangular portion represents a magnet. In FIGS. 11 and 12). The same). Therefore, the total number of magnets in the magnet array is 25. The first pitch p1 is twice as long as the second pitch p2. Reference numeral 24 denotes a liquid dripping prevention plate provided on the upper side of the magnet support block 25. When liquid is held in the dispensing tip 18 of the nozzle head 70, the dispensing tip 18 In order to receive liquid that may hang down from the mouth 18a, the magnet support block 25 is moved using the motor 27 or the like so as to be positioned below the dispensing tip 18. *

In step S10, the nozzle head 70 having the nozzles 12 ₁ to 12 ₁₂ mounted with the dispensing tip 18 holding the whole blood is moved in the Y-axis direction, and the first pitch of the first pitch is changed. It moves to the upper part of the said accommodating part 44a of the accommodating part group 40. Lysis 1 (enzyme) is accommodated in the accommodating portion 44a.

In step S11, by rotating the hexagonal shaft 85 provided on the nozzle head 70, the mouth portion 18a at the tip of the dispensing tip 18 is inserted into the accommodating portion 44a, and the pump 15 is driven. Then, the whole blood held in each of the dispensing tips 18 is mixed with the Lysis® 1 and stirred if necessary by repeating suction and discharge using the pump 15.

In step S12, the entire amount of the agitated liquid is sucked by the dispensing tip 18, and is accommodated in the reaction tube accommodated in the hole 46a set to 55 ° C. by the temperature control unit to perform incubation. Thereby, the protein contained in the whole blood is denatured. After a predetermined time has passed, while leaving the reaction solution in the reaction tube, the dispensing tip 18 is moved to the accommodating portion 44b by the Y-axis moving mechanism, and using the hexagonal shaft 85 and the pump 15, The entire amount of the liquid stored in the storage portion 44b is sucked, moved to the reaction tube using the Y-axis moving mechanism, the hexagonal shaft 85 and the pump 15, and discharged, and the total amount is used as a reaction solution. It discharges to the said accommodating part 44c. This destroys the protein and lowers the molecular weight. In addition, Lysis 収容 2 is accommodated in the accommodating portion 44b, and a binding buffer solution is accommodated in the accommodating portion 44c.

In step S13, the binding buffer solution as the separation / extraction solution housed in the housing portion 44c and the reaction solution are stirred to further dehydrate the solubilized protein, and the nucleic acid or fragment thereof is put into the solution. Disperse.

In step S14, the dispensing tip 18 and the pump 15 are used to suck the entire amount, and the hexagonal shaft 85 is used to raise the dispensing tip 18, and the reaction solution is removed using the Y-axis moving mechanism. Then, it moves to the upper side of the accommodating portion 44d, inserts the mouth portion 18a of the dispensing tip 18 into the accommodating portion 44d using the hexagonal shaft 85, and uses the pump 15 to insert the mouth portion 18a into the accommodating portion 44d. The first magnetic particle suspension contained in the magnetic particle suspension and the reaction solution are stirred, and a cation structure in which Na ⁺ ions are bonded to hydroxyl groups formed on the surfaces of the magnetic particles contained in the magnetic particle suspension. Is formed. Therefore, negatively charged DNA is captured by the magnetic particles.

In step S15, as shown in FIG. 9B or FIG. 11, the mouth portions 18a of the dispensing tips 18 arranged at the first pitch p1 are inserted into the accommodating portion 44d (FIG. 9 ( In b), the accommodation portion 44a is provided). However, when the magnet array provided in the magnet support block 25 is brought close to the narrow tube 18c of the dispensing tip 18, the first pitch p1 is the second pitch p2. The two magnets 22 and the adjusting magnets 22a and 22b on both sides are provided, so that one magnet 22 approaches the narrow tube 18c of the twelve dispensing tips 18. , Both sides will be sandwiched between two magnets, and the homogeneity will be high.

In step S16, with the magnetic field applied to each dispensing tip 18 in this manner, the liquid is discharged by repeating the suction and discharge by the pump 15, whereby the nucleic acid is applied to the inner wall of the thin tube 18c of the dispensing tip 18. The captured first magnetic particles are adsorbed and separated from the liquid. The nozzle head 70 is moved by the Y-axis moving mechanism in a state where the first magnetic particles are attracted to the inner wall of the thin tube 18c while the magnet 22 and the like are brought close to the thin tube 18c of the dispensing tip 18. It is moved along the Y-axis direction to the upper side of the next accommodating portion 44e. The cleaning liquid 1 (NaCl, SDS, isopropanol) is stored in the storage portion 44e.

In step S17, with the mouth portion 18a of the dispensing tip 18 inserted into the storage portion 44e, and with the magnet array having the magnets 22 separated from the dispensing tip 18 to remove the magnetic field, By driving the pump 15 and repeating the suction and discharge of the liquid, the magnetic particles are separated from the inner wall and stirred in the cleaning liquid 1 to remove the protein. After that, the magnet 22 is brought close to the thin tube 18c of the dispensing tip 18 again, and the magnetic particles are adsorbed on the inner wall of the thin tube, and the dispensing tip 18 is accommodated by using the Y-axis moving mechanism. Move to 44f. The cleaning liquid 2 is stored in the storage portion 44f.

In step S18, the dispensing tip 18 is lowered using the hexagon shaft 85, and the magnet 22 is separated from the dispensing tip 18 to remove the magnetic field, and is accommodated in the accommodating portion 44f. By repeatedly sucking and discharging the washing liquid 2 (isopropanol) using the pump 15, the magnetic particles are stirred in the liquid to remove NaCl and SDS, and the protein is washed. Thereafter, the magnetic tip (magnet 22 etc.) of the magnetic force device 20 is again brought close to the thin tube 18c of the dispensing tip 18 so that the magnetic particles are adsorbed on the inner wall of the thin tube, and the dispensing tip 18 is moved. After being raised by the hexagonal shaft 85, the Y-axis moving mechanism is used to move it to the accommodating portion 44g. Distilled water as a dissociation liquid is accommodated in the accommodating portion 44g.

In step S19, the dispensing tip 18 is lowered by the hexagonal shaft 85, and the washing liquid 2 is replaced with water by repeating suction and discharge of distilled water in a state where the magnetic force is applied to the dispensing tip 18. To remove. Thereafter, the magnetic particles are agitated by repeatedly aspirating and discharging the magnetic particles in distilled water as the dissociation liquid in a state in which the magnets 22 of the magnet row are all separated from the dispensing tip 18 and the magnetic force is removed. The nucleic acid or fragment thereof held by the particles is dissociated (eluted) from the magnetic particles into the liquid. Thereafter, the magnet 22 of the magnet array is brought close to the dispensing tip 18 to adsorb the magnetic particles to the inner wall, and the solution containing the dissociated nucleic acid remains in the storage portion 44g.

In step S20, the dispensing tip 18 holding magnetic particles is positioned above the accommodating portion 44h by the Y-axis moving mechanism. Distilled water is accommodated in the accommodating portion 44h.
With the hexagonal shaft 85, the dispensing tip 18 is lowered, the magnet 22 is separated from the dispensing tip 18 and the magnetic force is removed from the inside of the dispensing tip 18 to repeatedly suck and discharge distilled water. The magnetic particles are resuspended with the distilled water and discharged into the housing portion 44h to remove the magnetic particles.

In step S21, the dispensing tip 18 is raised by the hexagonal shaft 85, and is positioned above the accommodating portion 44i by the Y-axis moving mechanism. A second magnetic particle suspension is accommodated in the accommodating portion, and the magnetic particle suspension is attracted by the pump 15, and the magnet 22 is brought close to the thin tube 18 c of the dispensing tip 18. In this state, the second magnetic particle suspension is placed in a state where it is positioned above the reaction vessel 46b by the Y-axis moving mechanism and the dispensing tip 18 is lowered by the hexagonal shaft 85 and the magnet 22 is separated. Discharge.

In step S22, the dispensing tip 18 is lifted using the hexagonal shaft 85, and the dispensing tip is returned to the accommodation portion 44g using the Y-axis moving mechanism, and the nucleic acid contained in the accommodation portion 44g. In the same manner, the Y-axis moving mechanism moves the solution containing the solution up to the upper side of the reaction vessel 46b, and the dispensing tip 18 is lowered by the hexagonal shaft 85 to put the solution into the reaction vessel 46b. Discharge.

In step S23, the dispensing tip 18 is moved above the accommodating portion 44j by the Y-axis moving mechanism. The accommodating portion 44j accommodates a hybridization reagent such as DNA. The reagent is lowered by the hexagonal shaft 85 and sucked, and then lifted and moved to above the reaction vessel 46b by the Y-axis moving mechanism. The dispensing tip 18 is lowered by the hexagonal shaft 85 to remove the reagent. It discharges in the said reaction container 46b. These liquids are mixed and stirred by repeating suction and discharge using the pump 15 and incubated for a certain time. Thereby, the complementary nucleic acid or fragment thereof is bound to the nucleic acid or fragment thereof immobilized on the surface of the second magnetic particle.

In step S24, the magnet 22 of the magnetic device 20 is brought close to the thin tube 18c of the dispensing tip 18, and suction and discharge are repeated using the pump 15 to bind the target nucleic acid or a fragment thereof. The particles are adsorbed on the inner wall of the thin tube 18c, and the remaining liquid is discharged into the reaction vessel 46b. The dispensing tip 18 is raised using the hexagonal shaft 85.

In step S25, using the pitch conversion mechanism 60, the second magnetic particles attached to the nozzles 12 arranged at the first pitch p1 and capturing the target nucleic acid are adsorbed on the inner wall of the thin tube 18c. The interval between the chips 18 is reduced to the second pitch p2. At this time, when instructed by the control unit, the pitch conversion mechanism 60 drives the motor 65c to rotate the pulley 65a to travel the timing belt 65b for a predetermined distance and attach it to the timing belt 65b. By moving the movable plate 67 in the reference X coordinate position direction, the support plates 81 are moved along the X-axis direction, and the variable pitch between the nozzles is changed from the first pitch p1 to the second pitch p2. Perform conversion.

FIG. 9C and FIG. 12 show a state in which the variable pitch between the dispensing tips 18 is converted to the second pitch p2, and the first accommodation is performed using the Y-axis moving mechanism and the hexagonal shaft 85. A state in which the mouth portion 18a of the dispensing tip 18 is inserted from the portion group 40 into the one well row 52a of the microplate 51 of the second accommodating portion group 50 is shown. 12 nozzles are arranged in a second pitch p2 is closer to the stationary nozzle 12 _12, among the 23 pieces of magnets 22 of the magnet array, twelve magnets 22 and the outside of the magnets 22a and the magnet adjacent The 22 magnetic fields mainly affect each nozzle. The well row 52a contains the cleaning liquid 3, and the pump 15 is repeatedly used for suction and discharge to clean the second magnetic particles and remove impurities.

In step S26, the dispensing tip 18 is moved above the well row 52b by the Y-axis moving mechanism and lowered by the hexagonal shaft 85. In the well row 52b, a dissociation liquid is accommodated, and the magnet 22 is separated from the thin tube 18c of the dispensing tip 18, and the second magnetic particles are removed from the second magnetic particles by suction or temperature control by the pump 15. Dissociate the nucleic acid or fragment of interest. Thereafter, the magnet 22 of the magnet row is brought close to the thin tube 18c to apply a magnetic field, and the residual liquid is discharged to the well row 52b in a state where the magnetic particles are adsorbed on the inner wall of the thin tube 18c. Then, the cleaning liquid 4 stored in the well row 52c is sucked and discharged by the pump 15 while being moved to the upper side of the well row 52c by the Y-axis moving mechanism and the magnets 22 of the magnet row are separated from each other. By repeating the above, the second magnetic particles are resuspended and discharged to the well row 52c to be removed.

In step S27, the Y-axis moving mechanism and the hexagonal shaft 85 are used to return to the well row 52b to ascend and raise the liquid containing the target nucleic acid or fragment thereof, and transfer to the well row 52e for discharge. To do. Similarly, the Y-axis moving mechanism and the hexagonal shaft 85 are used to move to the well row 52d, the master mix is sucked, discharged to the well row 52e, and mixed and stirred.

In step S28, the microplate 51 is conveyed into the real-time PCR device 90 as a whole by the carriage 53, and temperature control based on the PCR method is performed. A real-time PCR process is performed to measure whether or not there is an error. For example, the target PCR product is detected using the nucleic acid labeled with a fluorescent substance in addition to the PCR primer containing the predetermined base sequence.

Subsequently, a magnetic particle reaction control device 100 using a variable pitch dispensing device according to a second embodiment of the present invention will be described based on FIGS. 13 to 16. Note that the same reference numerals as those in the first embodiment represent the same components, and thus the description thereof is omitted.
As shown in FIGS. 13 to 16, the magnetic particle reaction control device 100 using the variable pitch dispensing device according to the second embodiment uses the variable pitch dispensing device according to the first embodiment. In the magnetic particle reaction control apparatus 10, a nozzle head 700 is used instead of the nozzle head 70.

The nozzle head 700 has twelve nozzles 12 ₁ to 12 ₁₂ and the pitch conversion mechanism 60, similarly to the nozzle head 70, while the magnetic device 200 is replaced with the magnetic device 20 of the nozzle head 70. It is what you have. The magnetic device 200 is provided below the _twelve support plates 81 ₁ to 81 ₁₂ so that the magnetic poles are alternately reversed at the same pitch as the nozzles 12 ₁ to 12 ₁₂ along the X-axis direction. A variable pitch magnet array having twelve magnets 220 arranged so that S poles and N poles are alternately arranged on the side facing the nozzle, and twelve magnets 220 of the variable pitch magnet array are collectively A magnet pressing member 260 that can be moved forward and backward along the Y-axis direction and that extends in the X-axis direction that can be pressed against the thin tube 18c of the dispensing tip 18, and the magnet pressing member 260 attached to the magnet pressing member 260 is attached to the Y-direction. A shaft 260b (see FIGS. 14 and 15) extending in the Y-axis direction that can advance and retract along the axial direction, and the shaft 26 attached to a wall portion 72 attached to the substrate of the nozzle head 700. A linear motor 270 that drives the magnet array (magnet 220) along the Y-axis direction via b and the wall 72, and extends along the Y-axis direction. The magnet pressing member 260 extends in the Y-axis direction. And two guide rods 260a for guiding. Further, a detachable member that protrudes from the magnetic pole surface of the magnet 220 and allows the dispensing tip 18 to be detached from the nozzles 12 ₁ to 12 ₁₂ may be provided below the magnet row. Incidentally, it points nozzle 12 ₁₂ is stationary nozzle is similar to the first embodiment.

Further 12 of each magnet 220 as a variable pitch magnet rows are provided corresponding to 12 sheets of the support plate _{81 1-81 12,} the support plate _{81 1-81 12,} i.e., the nozzles ₁₂ 1 12 ₁₂ has the same variable pitch. The magnet 220 includes, as the magnet support members, an upper horizontal bar 263 attached to the lower side of the support plates 81 ₁ to 81 ₁₂ and two nodes provided at both ends of the upper horizontal bar 263. Two parallel links 261a and 261b that are respectively joined by a turning pair, and a lower side where the nodes at both ends are connected by a turning pair even at each node 264 provided at the lower end of the two links It has a rod 262 and is supported by the support plates 81 ₁ to 81 ₁₂ as a variable pitch magnet array. The magnet 220 is attached to one end of the lower horizontal bar 262 so as to protrude outward from the lower horizontal bar 262 on the lower side of the lower horizontal bar 262, and the magnet pressing member 260 is The lower horizontal bar 262 is provided so as to be in contact with the other end of the lower horizontal bar 262, and the lower horizontal bar 262 is always elastically biased so as to be separated from the dispensing tip 18.

According to the magnetic particle reaction control device 100 using the variable pitch dispensing device according to the second embodiment, the magnet 220 is provided for each dispensing tip 18 and is the same as the variable pitch of the nozzles 12 ₁ to 12 _12. The magnets 220 are arranged at a variable pitch. Therefore, in principle, the number of dispensing tips 18 and the number of necessary magnets 220 are the same, and the number of magnets can be reduced.

Each of the embodiments described above is specifically described for better understanding of the present invention, and does not limit other embodiments. Therefore, changes can be made without changing the gist of the invention. For example, as the nozzle, only the case of the dispensing tip mounted on the mounting nozzle has been described, but the present invention is not limited to this, and a nozzle to which the dispensing tip is not mounted may be used, and a pump as the suction / discharge mechanism Although only the case of using is described, it may be provided in the nozzle head itself. Also, a liquid and gas can be accommodated in the interior surrounded by the wall surface, and a deformed wall surface capable of predetermined deformation of the wall surface is formed in a part of the wall surface, and the deformed wall surface communicates with the housing portion. A bellows type dispensing tip having a mouth part through which the liquid sucked and discharged by the internal expansion and contraction due to the deformation can flow in and out may be used together with a mechanism for deforming the dispensing tip.

Furthermore, in the above description, only the Y-axis movement mechanism has been described as the nozzle head movement mechanism, but an X-axis movement mechanism or a Z-axis movement mechanism may be used. Furthermore, instead of the nozzle head, the first housing portion group, the second housing portion group, and the like may be movable along the Y-axis direction or the X-axis direction, or both may be movable.

Further, in the above description, only the case where 12 links are used as the pitch conversion mechanism has been described. However, the present invention is not limited to this case. For example, the nozzles for mounting each dispensing tip arranged in a single row are used. And the cylinder are supported by the support portion so as to be displaceable along the row direction at each arrangement position. In addition, a cross-shaped member in which a plurality of rod-shaped members having the same shape are connected to each other so as to be rotatable at the center fulcrum, and the other cross-shaped member is provided at each fulcrum provided at each tip of the two rod-shaped members And four rod-shaped members to form one rhombus frame, and each fulcrum between the centers has a number of nozzles for mounting, etc. (12 from the start fulcrum to the end fulcrum, 11 of the rhombus frames are formed such that either one of the start fulcrum or the end fulcrum is fixed to the support portion, and the other is movable in the row direction). An expansion / contraction connection mechanism combined so as to be expandable / contractible in the row direction, and a pitch variable mechanism in which nozzles and cylinders for mounting the dispensing tips are connected to the central fulcrums. Alternatively, cams are provided between the adjacent dispensing tips or the mounting nozzles and cylinders, and the adjacent dispensing tips are always urged in the direction of being narrowed by a spring, and the cams are simultaneously moved by a rotating mechanism. It may be a pitch conversion mechanism that converts the pitch by rotating it to the right.

In the above description, only 12 nozzles, 12 cartridge containers, 96-well microplates, and 25 or 12 magnets are used. However, the present invention is not limited to these cases. Depending on the number of nozzles (for example, 4, 8, 10, etc.), various numbers of cartridge containers, microplates with a number of wells, and other numbers of magnets can be handled. Moreover, although the nucleic acid extraction process and the real-time PCR process have been described as process examples, the process is not limited thereto. Note that “row” and “column” are convenient and can be used interchangeably. Further, spaces such as “upper”, “lower”, “inner”, “outer”, “row direction”, “column direction”, “X axis”, “Y axis”, “Z axis”, etc. within the present application. The representation is for illustration only and is not limited to a specific spatial orientation or placement of the structure.

Magnetic particle reaction control device using the variable pitch dispensing device according to the present invention and its control method, fields that require processing of various solutions, for example, the agricultural field such as industrial field, food, agriculture, fishery processing, It relates to all fields such as pharmaceutical field, hygiene, insurance, immunity, disease, genetic field, medical field, chemistry or biology field. The present invention is particularly effective when a series of processes using a large number of reagents and substances are performed in parallel in a predetermined order for a large number of objects. *

10,100 Magnetic particle reaction control device using variable pitch dispensing device 12 ₁ to 12 ₁₂ nozzles 12 ₁₂ stationary nozzles
18 Dispensing tip (nozzle tip)
20,200 Magnetic device 21 Detachable member 22,220 Magnet (magnet array)
24 Liquid dripping prevention plate 25 Magnet support block (magnet support member)
30 3rd accommodating part group 40 1st accommodating part group 50 2nd accommodating part group 60 Pitch conversion mechanism 70,700 Nozzle head 80 Z-axis independent moving mechanism (independent nozzle moving mechanism)
90 Real-time PCR processing equipment

Claims (15)

A plurality of nozzles arranged at a variable pitch in at least one row having a tip portion capable of sucking and discharging liquid, and a pitch capable of converting the nozzles into at least a first pitch and a second pitch according to instructions A nozzle head having a conversion mechanism, and a first portion having a plurality of receiving portions that can be inserted all at once at the first pitch and arranged in one or more rows along the row direction at the first pitch. A second accommodating portion group having a plurality of accommodating portions arranged in one or more rows along the row direction at a second pitch smaller than the first pitch, and the nozzle A moving mechanism that allows the head or the nozzle to move relative to the first housing portion group and the second housing portion group, and at least the first pitch or the second pitch. in front A magnetic device capable of simultaneously applying and removing a magnetic field in the tip portion of the nozzle, and arranged along the row direction in either the first housing unit group or the second housing unit group A magnetic particle reaction control device using a variable pitch dispensing device in which a suspension of magnetic particles is accommodated in the accommodating portion. The magnetic device is provided in the nozzle head, and one or a pair of magnets provided corresponding to the nozzles is at least along the row direction at the first pitch or the second pitch. The magnet rows arranged in a row and the magnets corresponding to the tip portions of the nozzles when the variable pitch of the nozzles is set to the first pitch or the second pitch are collectively shown. A magnetic particle reaction control device using the variable pitch dispensing device according to claim 1, further comprising: a magnet row moving mechanism that moves the magnet row so that the magnet row can be contacted and separated. In the magnet row, the one or one set of magnets corresponding to the nozzles are arranged in at least one row along the row direction at the first pitch and the second pitch, and the magnet row moving mechanism The magnets are configured so that the magnets corresponding to the tip portions of the nozzles can be simultaneously contacted and separated when the variable pitch of the nozzles is set to the first pitch and the second pitch. The magnetic particle reaction control device according to claim 2, wherein a variable pitch dispensing device that enables movement of the row is used. The magnetic device is provided in the nozzle head, and one or a pair of magnets provided corresponding to the nozzles are variablely arranged in at least one row at the same variable pitch as the variable pitch of the nozzles. A pitch magnet row; and a magnet row moving mechanism for moving the magnet row so that the arranged magnets can be simultaneously contacted and separated from the corresponding tip portions of the nozzles. The magnetic particle reaction control device using the variable pitch dispensing device according to claim 1, wherein the magnet row is pitch-converted by the pitch conversion mechanism together with the nozzle. An independent nozzle moving mechanism capable of raising and lowering the plurality of nozzles independently of each other with respect to the nozzle head; and the pitch converting mechanism is capable of converting the variable pitch of the nozzles to a third pitch. , Having a third housing portion group having a plurality of housing portions arranged in one or more rows along the row direction at the third pitch, and the third housing portion group includes a sampling portion The magnetic particle reaction control apparatus using the variable pitch dispensing apparatus according to claim 1, wherein sample storage portions capable of storing the prepared sample solution are arranged in at least one row along the row direction. The third pitch is larger than the first pitch, and the first storage section group includes at least one storage section that stores at least one kind of reagent solution for reacting the sample solution along the column direction. The storage units arranged in one row and containing the magnetic particle suspension are arranged in at least one row along the row direction, and the temperature-controllable reaction vessels are arranged in at least one row along the row direction. 6. The magnetic particle reaction control using the variable pitch dispensing device according to claim 5, wherein the second storage section group is configured such that the wells for light measurement are arranged in at least one row along the row direction. apparatus. Each nozzle includes a mounting nozzle and a tip portion that is detachably mounted on the mounting nozzle, and the variable pitch of the nozzle is set to the first pitch, the second pitch, or the third pitch. In this case, the tip portion is accommodated at the first pitch, the second pitch, or the third pitch in a state where the tip portion can be attached to the mounting nozzle by lowering the mounting nozzle. The magnetic part using the variable pitch dispensing device according to any one of claims 1 to 6, further comprising: a tip portion storage portion group in which the tip portion storage portions to be arranged are arranged in at least one row along the row direction. Particle reaction control device. The nozzle includes a stationary nozzle that is stationary when the variable pitch of the nozzle by the pitch variable mechanism is converted into at least a first pitch and a second pitch, and the first accommodating portion group and the second The accommodation unit group is arranged along the linear movement path of the stationary nozzle by the movement mechanism, the stationary nozzle corresponding accommodation unit into which the tip of the stationary nozzle is to be inserted, and the magnet array The magnetic particle reaction control device using the variable pitch dispensing device according to any one of claims 2 to 7, wherein each magnet is arranged with reference to a position corresponding to the stationary nozzle. The variable pitch of the nozzle of the nozzle head in which a plurality of nozzles having tip portions capable of sucking and discharging liquid are arranged in one or a plurality of rows at a variable pitch that can be converted by an instruction is set to the first pitch. A first pitch setting step to set;
The nozzle head or the nozzle is moved relative to the first housing portion group having a plurality of housing portions arranged in one or more rows along the row direction at the first pitch, and suction is performed. Or a first suction / discharge step for discharging;
The nozzle head or the nozzle is moved relative to the first accommodating portion group, and the tip portion of the nozzle is extracted from the accommodating portion, and the variable pitch of the nozzle is set to be larger than the first pitch. A second pitch setting step for setting a small second pitch;
A second for performing suction or discharge by moving the nozzle head or the nozzle relative to a second housing portion group having a plurality of housing portions arranged in one or a plurality of rows at a second pitch. And a suction discharge process of
Magnetic particle suspensions are accommodated in the accommodating portions arranged in at least one row along the row direction in either the first accommodating portion group or the second accommodating portion group, and the first suction discharge A magnetic field is applied to the tip of each nozzle arranged in at least one row in the row direction at the first pitch or the second pitch at the time of suction or discharge in the step or the second suction / discharge step. By the magnetic field separation step of adsorbing and separating the magnetic particles on the inner wall of the tip, and removing the magnetic field from the tip of each nozzle during the suction or discharge, to the inner wall of the tip A magnetic particle reaction control method using a variable pitch dispensing device further comprising a resuspension step of separating the adsorbed magnetic particles from the inner wall and resuspending them in a liquid. In the magnetic field separation step, at the time of the suction or discharge in the first suction / discharge step or the second suction / discharge step, one or a set of magnets corresponding to each nozzle is at least the first pitch or A magnetic field is obtained by moving a magnet row arranged in the row direction in at least one row at a second pitch and bringing the one or one set of magnets close to the tip of each corresponding nozzle all at once. The resuspension step is performed by moving the magnet row during the suction or discharge through the tip of the nozzle to move from the tip of the nozzle. The magnetic particle reaction control method using the variable pitch dispensing device according to claim 9, wherein the corresponding one or one set of magnets are separated at a time. At the time of the suction or discharge in the first suction / discharge step and the second suction / discharge step, at least one magnet or one set of magnets corresponding to each nozzle is at least at the first pitch and the second pitch. 11. A magnetic field is applied to each tip by moving a magnet row arranged in a row and bringing the one or one set of magnets close to the tip of each corresponding nozzle all at once. Magnetic particle reaction control method using a variable pitch dispensing apparatus. The magnetic field separation step is provided in the nozzle head at the time of the suction or discharge in the first suction / discharge step or the second suction / discharge step. One set of magnets is moved at least in one row at a variable pitch that is the same as the variable pitch of the nozzles, and the one or one set of magnets is moved to the tip of each corresponding nozzle. A magnetic field is applied to the inside of each tip by approaching all at once, and the resuspension step moves the magnet row during the suction or discharge through the tip of the nozzle, The magnetic particle reaction control method using the variable pitch dispensing device according to claim 9, wherein the corresponding one or one set of magnets are separated from the tip of each nozzle at once. Prior to the first pitch setting step, a third pitch setting step of setting the variable pitch of the nozzle head to a third pitch larger than the first pitch, and the third pitch at the third pitch The nozzle head or the nozzle is relatively moved with respect to a third housing portion group having a plurality of housing portions arranged in one or a plurality of rows in the row direction, and the nozzles are mutually independent. And a third suction / discharge step for sucking the collected sample solution stored in the storage section arranged along the row direction of the third storage section group while being lifted and lowered, and the first suction In the ejection step, the storage unit of the first storage unit group and the magnetic particle suspension arranged in at least one row along the column direction in which at least one reagent solution for reacting the sample solution is stored. In the direction of the row where the turbid liquid is contained A step of performing suction and discharge with respect to the storage portions of the first storage portion group arranged in at least one row, and after the second suction and discharge step, at least one row along the column direction. A magnet using the variable pitch dispensing device according to claim 9, further comprising: a light measuring step of measuring light with respect to the light measuring wells of the second housing group arranged in a shape. Particle reaction control method. The nozzle includes a mounting nozzle and a tip portion that is detachably mounted on the mounting nozzle, and the variable of the mounting nozzle of the nozzle head is performed before or in the first pitch setting step. The pitch is set to a first pitch, a second pitch, or a third pitch larger than the first pitch, and the nozzle head or the mounting nozzle is mounted in the state in which the tip portion can be mounted. The tip portion is moved to the tip portion accommodating portion group accommodated in the plurality of accommodating portions arranged at the pitch, the second pitch, or the third pitch, and the tip portion is moved to the attachment nozzle by lowering the attachment nozzle. A magnetic particle reaction control method using the variable pitch dispensing device according to any one of claims 9 to 13, further comprising a step of attaching the step. In the first pitch setting step and the second pitch setting step, among the plurality of nozzles arranged in at least one row along the row direction provided in the nozzle head, the nozzle is fixed during pitch conversion. The stationary nozzles in the first housing part group and the second housing part group in such a manner that the movement path in the movement in the first suction / discharge process and the second suction / discharge process is linear. The stationary nozzle corresponding accommodating portion into which the stationary nozzle is to be inserted is arranged along the movement path, and the magnet is arranged in the separation and extraction step with reference to the stationary nozzle. A magnetic particle reaction control method using the variable pitch dispensing device according to claim 14.

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