WO2001070403A1 - Collection vessel for separating biological fluid constituents - Google Patents

Abstract

The invention relates to a collection vessel (1) for fluids (2) comprising a vessel interior (3) which is at least partially enclosed by a vessel lateral surface (6) and, optionally, by a vessel bottom (10) that form(s) a vessel inner wall (5), and which is accessible via at least one vessel opening (4). In the vessel interior (5), at least one reagent or reagent mixture (20), with which at least individual constituents of the fluid (2) or cleavage products of these constituents can be separated out and/or immobilized and/or stabilized, is placed on a support (21). Said support (21) can be formed by the vessel inner wall (5) and/or by a closing device (11) for the vessel opening (4), said closing device being preferably provided with a septum (12), or the support can be formed by the reagent or reagent mixture (20).

Description

 COLLECTIBLE FOR THE SEPARATION OF BIOLOGICAL LIQUID COMPONENTS

The invention relates to a collecting vessel for liquids of biological origin or biological matrices containing according to the features in the preamble of claim 1, a carrier for reagents or reagent mixtures according to the features in the preamble of

Claim 27, an analysis kit for analyzing liquids of biological origin or biological matrices containing according to the features in the preamble of claim 45 and a method for separating and / or immobilizing and / or stabilizing components or fission products of such a liquid according to the features in the claim 51st

In the field of laboratory diagnostics, the past has been shaped by designing analytical methods and analysis devices for the analysis of liquids of biological origin, such as blood, more and more efficiently, not least in order to lower the detection limit for certain blood components. However, it is still necessary to analyze complex blood components such as genetic information, e.g. deoxy- and ribonucleic acid (DNA, RNA), a multitude of different preparation steps is required. One reason for this is that the DNA or RNA are not stable or are only stable to a limited extent and are therefore subject to a continuous degradation process from the time the blood is drawn, triggered by nucleases,

Enzymes or the like, which are contained in cells, and thus the result of the analysis is falsified by unwanted fission products. A replication of the physiological control, with which the effect of cellular and extracellular nucleases does not come into play as long as the cells are in their normal environment, has so far not been successful. The withdrawal of blood leads to more or less strong changes in the cells

Nucleic acids, so that long-term storage of blood leads to aging or destruction of the cell. This naturally also leads to problems when examining viruses contained in the blood via their genetic coding.

One of the common ways today to keep blood unchanged over a long period of time is to lower the storage temperature, for example with the help of liquid nitrogen. Another possibility is to add certain additives to the blood or generally a biological fluid. These additives should be selected so that they do not interfere with a subsequent analysis. A method is known from EP 0 818 542 A1, with the aid of which a guanidini salt in powder form the nucleases are to be inhibited.

DE 195 19 886 AI describes a blood collection device which can be closed by a puncturable stopper. This stopper is designed such that a vessel with an additive contained therein projects into a recess, this vessel having a bottom which can also be punctured but cannot be closed again. A cannula piercing the stopper penetrates the vessel with the additive and the further advancement of the cannula pierces the bottom thereof. This makes it possible to bring the additive into contact with the blood while the blood is being drawn. With the aid of this technique, the disadvantage which arises from the use of polyethylene terephthalate tubes, namely its water vapor permeability and thus the impairment of the additive present in the blood collection device, for example by clumping, is to be eliminated.

A disadvantage of both the method according to EP-AI and the blood collection device according to DE-Al is that in principle only a single process step in the chain of preparation steps necessary for the analysis of blood constituents is covered, namely in the case of the EP -AI the stabilization of the nucleic acid and, in the case of DE-Al, the addition of an additive which prevents blood clotting or accelerates blood clotting. In addition, DE-A1 has the disadvantage that there is no defined addition of the additive, since an unspecified residue always remains in the vessel.

EP 0 875 202 A2 shows another way of introducing an additive into a blood sample or into a part of a blood sample. For this purpose, this includes blood collection tubes in the lower

Part of a matrix of fiber material, which is impregnated with a solution of an aqueous additive. To protect the additive, this blood collection tube also contains a water-immiscible liquid, which has a specific weight greater than 1.07, and the aqueous additive can thus be encapsulated. A device is also included in the blood collection tube, which allows a clean separation of the phases after filling the tube and centrifuging. With the help of this system, a coagulation reagent is to be added to the blood sample and in turn only a single step of the preparation phase for the detection of individual components of the blood can be covered.

The invention has for its object to provide a way to analyze Simplify liquids of biological origin or biological matrices.

This object of the invention is achieved by the features in the characterizing part of claim 1. It is advantageous that the user now has a collecting vessel, e.g. for blood collection, which can be used not only to absorb the liquid, but at the same time, preferably with the introduction of the liquid into this collecting vessel, a separation of the liquid or fission products of this liquid and, if appropriate, an at least partial analysis of individual components this liquid takes place. It can e.g. the blood draw, in particular the time until an analysis result is available, can be shortened and, if necessary, carried out by the user without the involvement of a laboratory.

An embodiment according to claim 2 is also advantageous, since it makes the genetic information available more quickly and, if necessary, nucleic acids can be stored in the matrix over a longer period of time, as is advantageous, for example, for the investigation of criminal cases.

An embodiment according to claim 3 is also advantageous, since it enables an indirect, qualitative statement to be made about the presence or absence of nucleic acids or nucleic acid mixtures.

Another advantage is an embodiment according to claim 4, according to which a disruption of the cells is already possible during the filling process of the collecting vessel.

However, an embodiment according to claim 5 is also advantageous, since it enables the substance to be analyzed to be bound to a solid phase and the latter to be separated from the matrix together with the substance to be analyzed. Furthermore, it is advantageous in this embodiment variant that a possibility can be created with which reagents can be made available in the liquid over a longer period of time depending on dissolving behavior.

With the development according to claim 6, the advantage can be achieved that the collected liquids can be mixed with the reagent or reagent mixture without great effort. Furthermore, an embodiment according to claim 7 is possible, with the aid of which substances to be analyzed, which are not accessible or can only be accessed with great effort, can be examined.

By forming the reagent or reagent mixture according to claim 8, a division of the constituents of the liquid 2 into several phases and thus a simpler separation of the substances to be examined can be achieved.

It is also advantageous to design the reagent or reagent mixture in accordance with claim 9, since by using at least one of these chaotropic salts

Examination of genetic material, in particular the stability of nucleic acids in the matrix, can be improved.

However, a further development according to claim 10 is also advantageous, since it effectively provides a collecting vessel with a reagent or reagent mixture of a certain concentration

A separation according to density gradients can be made available.

However, an embodiment of the collecting vessel according to claim 11 is also advantageous, according to which a separation of sensitive substances from the matrix and thus a stable position of the liquid to be examined is possible.

Furthermore, an embodiment variant according to claim 12 is possible, according to which a precipitation and thus a simple separation of the substance to be examined is possible, for example by filtration, decanting, etc. from the matrix.

A configuration according to claim 13 is also advantageous, since it can facilitate the analysis of hydrophobic substances.

According to an embodiment according to claim 14, the concentration of individual constituents of the liquids to be examined and thus, in the end, a faster analysis is possible, since these steps are omitted in the subsequent laboratory tests.

Through the development according to claim 15, it is possible to design the reagent or reagent mixture so that it can be used both for analysis and for the physical separation of the constituents of the liquid. According to the development according to claim 16, the advantage can be achieved that a larger surface is available for the reaction of the substance to be analyzed with the reagent or reagent mixture.

In accordance with the configuration according to claim 17, it is possible to separate the carrier from the rest of the liquid either together with the substance to be analyzed or with the matrix.

An advantageous surface enlargement of the carrier can be achieved by the configurations according to claims 18 and 19.

According to the embodiment according to claim 20, a separation by molecular weight and thus a corresponding fractionation of the substances to be analyzed is possible.

Through the development according to claim 21, the carrier can be easily adapted to a wide variety of requirements.

However, a further development according to claim 22 is also advantageous, since it enables a simple separation of the carrier from the matrix.

By designing the collecting vessel according to claim 23, any liquids can be divided and separated into several phases, regardless of whether it is clear before the use of the collecting vessel whether the substance to be analyzed is present in the heavier or lighter phase.

A further development according to claim 24 is also advantageous, whereby a medium-heavy phase can be separated from a light and a heavy phase without the latter having to be removed from the vessel.

However, an embodiment according to claim 25 is also possible, since it enables the components of the liquid to be examined to be divided into several phases using laboratory aids.

It is also possible to implement a variant according to claim 26, with the individual Components of the liquid to be analyzed are selectively possible from the matrix without the aid of further auxiliary substances.

The object of the invention is also achieved by a carrier according to the features in the characterizing part of claim 27. The advantage here is that both the carrier and the reagent or reagent mixture can be easily adapted to the problem posed and such carriers can also be used universally in any vessel. In particular, this configuration of the carrier allows it to be separated from the liquid to be analyzed in a simple manner, for example with the aid of mechanical means.

It is advantageous if, according to claim 28, the surface of the carrier body is coated with the reagent or reagent mixture, since a subsequent separation of the used reagent or reagent mixture or the adhering substance to be analyzed from the carrier is possible and thus the latter under certain circumstances can be reused after appropriate preparation.

Embodiments according to claims 29 to 31 are advantageous, whereby a correspondingly enlarged surface can be made available for the reaction of the substance to be analyzed.

Due to the development according to claim 32, the carrier according to the invention can be introduced into and removed from a liquid to be analyzed in a simple manner.

An embodiment according to claim 33 is also advantageous, as a result of which a selective use of polarizable or polar substances is possible.

A certain cohesion of the individual particles and thus their easier removal from the liquid to be analyzed is made possible by the configurations according to claims 34 to 36, wherein at the same time this carrier can, for example, fill the entire cross-section of a vessel receiving it without the passage of the liquid is impaired too much, at the same time an increase in surface area can be achieved. A configuration according to claim 37 is also advantageous, according to which, in particular, an adaptation of the available carrier body surface to elongated analysis vessels is possible.

Furthermore, it is possible to provide the carrier with a closure device according to claim 38, so that the carrier can simultaneously fulfill the closure function after being introduced into a corresponding vessel.

A further development in accordance with claim 39 is advantageous, according to which a tight closure of the corresponding collecting vessel can be achieved and thus an intimate one

Mixing the liquid to be analyzed with the carrier or the components of the carrier is possible without spilling the liquid.

A configuration according to claim 40 is also possible, since it can be achieved that a vessel closed with such a carrier can be filled without this carrier having to be removed from the vessel.

Embodiments according to claims 41 and 42 are also advantageous, since this enables the carrier to supply further reagents or washing liquids in a simple manner.

However, a further development according to claim 43 is also advantageous, since it enables the carrier to be adapted to any cross-section.

Due to the configuration according to claim 44, the carrier can be easily adapted to a wide variety of uses.

The object of the invention is further achieved by an analysis kit according to the features in the characterizing part of claim 45. It is advantageous here that the user can be provided with a coordinated system for analyzing liquids, with which a rapid examination can be made possible while simultaneously reducing the necessary steps.

By arranging further collecting vessels according to claim 46, the advantage can be achieved that within a closed system several work steps for Analysis of liquids can be carried out and it is also possible in this way to achieve a simple separation of components of the liquids.

In this case, an embodiment according to claim 47 is advantageous since, after a plurality of collecting vessels have been separated from one another, the individual collecting vessels are automatically sealed.

A further simplification of the analysis can be achieved by compiling the analysis kit according to claims 48 to 50.

Finally, the object of the invention is also achieved by a method according to the features of the characterizing part of claim 51. It is advantageous that certain analysis steps or separation operations can be carried out while the liquid to be analyzed is being introduced, and at the same time stabilization for longer storage before processing of the analysis products becomes possible.

An embodiment in accordance with claim 52 is advantageous, since in this way it is possible to separate different phases of the liquid to be analyzed, so that subsequent, unintentional mixing or mutual influencing of the two phases can be prevented.

Finally, however, further developments are also possible according to claims 53 and 54, with which reactions to possible stability problems of reaction products in the matrix can be reacted to in a simple manner.

The invention is explained in more detail below on the basis of the exemplary embodiments illustrated in the drawings, with all representations being carried out in a greatly simplified manner.

Show it:

1 shows a collecting vessel according to the invention with a coating of a reactive component applied to the inner wall;

2 shows a collecting vessel according to the invention with an inserted carrier, which is provided with a reactive component; 3 shows another embodiment variant of the collecting vessel according to the invention with the carrier inserted;

4 shows a collecting vessel according to the invention with coated carrier particles arranged in the interior of the collecting vessel;

5 shows a variant of the arrangement of the carrier particles in the collecting vessel;

6 shows a carrier for introducing the reactive component into the collecting vessel;

7 shows another embodiment variant of the carrier according to the invention;

8 shows a collecting vessel according to the invention with a liquid reactive component;

9 shows a collecting vessel with a support for the reactive component that can be screwed onto it;

10 shows an analysis kit consisting of a collecting vessel and a plurality of carriers which have been put on;

11 shows an analysis kit consisting of a plurality of collecting vessels which communicate with one another via cannulas;

12 shows a collecting vessel according to the invention with a separation device used for better phase separation during or after centrifugation;

13 shows a collecting vessel according to the invention with a cap, in which a carrier with the reactive component is contained;

14 shows the carrier for the reactive component as part of a cannula;

15 shows a collecting vessel with a carrier for the reactive component arranged on a closure device for the collecting vessel;

16 shows a two-vessel system, with a reactive component container on the inner vessel. Layering is appropriate;

17 shows a further embodiment variant of the collecting vessel according to the invention in a simplified, schematic representation;

18 shows an embodiment variant of the collecting vessel for multi-component analysis;

FIG. 19 shows the closure device for the collecting vessel according to FIG. 18 in a schematically simplified plan view.

To begin with, it should be noted that in the differently described embodiments, the same parts are provided with the same reference numerals or the same component names, and the disclosures contained in the entire description can be applied analogously to the same parts with the same reference numerals or the same component names. The location information selected in the description, e.g. above, below, laterally etc. refer to the figure described and illustrated immediately and are to be transferred to the new position in the event of a change of position. Furthermore, individual features or combinations of features from the different exemplary embodiments shown and described can also represent independent, inventive or inventive solutions.

In Fig. 1, a collecting vessel 1 according to the invention for liquids 2 of biological origin or biological matrices is shown in a diagram in section. A vessel interior 3 is accessible through at least one vessel opening 4. The interior 3 of the vessel is delimited by an inner wall 5 of the vessel, which consists of a vessel jacket 6 with two vessel jacket end faces 8, 9 arranged opposite one another with respect to a vessel jacket center axis 7, and optionally, as shown in FIG. 1, a vessel bottom with the vessel jacket end face 9 10 can be connected in one piece, and at least one closure device 11 closing the vessel opening 4 is formed. This collecting vessel 1 can be designed, for example, as a blood collection tube known from the prior art. However, it is also possible to use the collecting vessel 1 to hold other liquids 2.

The collecting vessel 1 can, for example, be made of a preferably thermoplastic synthetic material. Substance, for example polyethylene terephthalate (PET), polypropylene (PP), polyvinyl chloride (PVC), polystyrene (PS) or the like, glass, metal, ceramic or the like., Or several of these materials can also form the collecting tank 1.

The closure device 11 is preferably designed so that a perforable therein

Septum 12 is held. For this purpose, the closure device 11 can have an outer jacket 13, which in turn can be made of plastic. The closure device 11 can e.g. be designed as a screw cap, plug cap or the like and, in order to enable perforation of the septum 12, have a bore 14 in the plane parallel to the end faces 8 of the vessel jacket. Otherwise, the closure device 1 1 from the prior art

Technically known closure devices for example for blood collection tubes correspond.

It is advantageous if the outer jacket 13 overlaps the vessel jacket 6 in the direction parallel to the vessel jacket center axis 7, at least in regions, since when the filled jacket is opened

Collecting vessel 1, the risk of inadvertent splashing out of the liquid 2 can be largely prevented.

The attachment of the closure device 11 to the collecting vessel 1, i.e. on the vessel jacket 6, e.g. Via a thread 16 arranged on an outer jacket inner side 15 of the closure device 11, indicated by dashed lines in FIG. 1. For this purpose, a corresponding web 17 can be arranged on the vessel jacket 6, for example in the region of the vessel jacket end face 8, so that engagement of this web 17 in the thread 16 is possible or it is also possible to provide an outer wall 18 of the vessel jacket with a corresponding thread ,

It is also possible, in particular if the closure device 11 is designed as a plug closure, that the closure device 11 is attached solely via adhesive forces between the septum 17 and the inner wall 5 of the vessel. These adhesive forces can of course have a supporting effect in the variant with the thread 16.

Of course, it is also possible for the closure device 11 to be formed solely by the septum 12, or other closure devices 11 which, for example, have no bore 14, can also be used. Perhaps the most suitable material for the septum 12 are perforatable materials, for example elastomers, for example rubber or the like. This septum 12 is intended to fulfill the task that the perforation, which is caused by the puncture of the septum 12 with a cannula 19 or a corresponding other suitable device, is closed again after the cannula 19 has been pulled out so that the liquid can escape 2 is prevented from the collecting vessel 1. Contamination of the user of the collecting vessel 1, for example with bacteria, viruses or the like, which can be contained in the liquid 2, is thus to be avoided, as by the above-mentioned design of the closure device 11.

Preferably, in the interior 3 of the vessel, there is a 1013 mbar at 298.15 K, lower pressure prevailing. It can thus be achieved that the liquid 2 is automatically brought into the interior 3 of the vessel as a result of this pressure drop.

The pressure in the interior can be set so that a predefined volume of liquid is introduced into the interior 3 of the vessel.

As shown in FIG. 1, the vessel bottom 10 can be semicircular or it is also possible to choose other embodiments.

According to the invention, at least one reagent or reagent mixture 20 accessible through the vessel opening 4 is arranged in the interior 3 of the vessel, with which at least individual components of the liquid 2 or the biological matrix or fission products of these components can be separated and / or immobilized and / or stabilized. The arrangement can be such that the inner wall 5 of the vessel is coated with this reagent or reagent mixture 20. This coating can be applied using all conventional, suitable techniques which are known from the prior art, for example it is possible to suspend the reagent or reagent mixture 20 in an appropriate solvent, which should have as high a vapor pressure as possible Collection vessel 1 is introduced, whereupon excess suspension is removed again and the

Drying of the reagent or reagent mixture 20, the solvent is evaporated, if necessary with an increase in temperature, so that in the end the reagent or reagent mixture 20 is in the solid state.

Of course, known methods of spray technology for coating the Collection vessel 1 can be used with the suspension or the reagent or reagent mixture 20.

The reagent or reagent mixture 20 can, for example, be selected or composed such that it is selective for nucleic acids or nucleic acid mixtures. The selectivity can be designed both for individual nucleic acids, for example deoxyribonucleic acids (DNA) or ribonucleic acids (RNA). In this way it is possible to bind nucleic acids of various origins to this reagent or reagent mixture 20, so that after removal of the remaining liquid matrix these nucleic acids can then be eluted from the reagent or reagent mixture 20 or the reaction products formed can be processed and are directly available for further analysis. In this way, the advantage can be achieved that the substance to be analyzed or the substance mixture can be prepared so far in essentially one step that its detection is possible either directly or with the interposition of only a few additional preparation steps (e.g. elution of the analyte to be analyzed Substance). For this purpose, the reagent or reagent mixture 20 should be selected or composed such that, in the case of the examination of nucleic acids in the blood, stabilization and / or lysing takes place during or shortly after the blood withdrawal and the subsequent immobilization of the nucleic acids takes place.

Lysing is necessary because blood is usually a mixture of cells, proteins, metabolites, etc. The cells in turn consist of eritrocytes, platelets and leukocytes, only the latter containing the nucleic acids. In order to bring these nucleic acids to a possible reaction, it is necessary to lyse the cells.

For a qualitative analysis it is further possible that the reagent or reagent mixture contains a substance which reacts selectively for a certain nucleic acid, for example in order to make the nucleic acid accessible for subsequent fluorescence detection.

There is also the possibility that a controlled degradation of the nucleic acid takes place with the aid of the reagent or reagent mixture 20 and that the reagent or reagent mixture 20 reacts selectively to these nucleic acid components, for example mononucleotides, oligonucleotides or the like. Furthermore, it is possible that, in the event that the liquid contains 2 constituents which react specifically with the particular nucleic acids, the reagent or reagent mixture 20 is selected selectively for these substances. Such nucleic acid Degrading components can be, for example, proteins, metabolites, nucleases, enzymes or the like.

In a modification of this embodiment variant, it is possible to use materials for the collecting vessel 1 which at least partially form the reagent or reagent mixture 20. For example, it has long been known that glass surfaces can selectively bind nucleic acids. This principle can also be used for plastics if they are modified accordingly. For example, it is possible to use polymers which have terminal, charged or chargeable groups, for example by acid treatments. Examples include amino groups or hydroxyl groups, in which amino groups, e.g. Diethylamino groups, through an appropriate acid treatment step the nitrogen can be positively charged and thus the binding of, for example, the DNA via the phosphate group, i.e. in particular an oxygen of this phosphate group becomes possible. Likewise, of course, nitrile polymers or copolymers or the like can be used, but care must always be taken that these

Have polarizable or polarized, preferably unbranched substituents corresponding to polymers. With the aid of this design of the collecting vessel 1, it is possible in a simple manner if an additional reagent or reagent mixture 20 is present in the collecting vessel 1, which takes over the lysis and / or stabilization of blood samples, in particular nucleic acids, preferably separating them quantitatively. Since these are selectively held by the wall of the collecting vessel 1, it is also possible to pour off the excess portion of the blood sample, so that the collecting vessel 1 can subsequently be used for washing and for further processing of the nucleic acids.

Another example of such an embodiment variant of the collecting vessel 1 is the use of a plastic with carboxy groups, which enables the attachment of oligonucleotides. The corresponding lysis of the cells is of course also required in this embodiment variant.

Another embodiment variant of the collecting vessel 1 according to the invention is shown in FIG. In contrast to the above variant, the reagent or reagent mixture 20 is not arranged as a coating on the inner wall 5 of the vessel, but is instead located on its own carrier 21. This carrier 21 can be connected to the vessel 1, for example by gluing to the inner wall 5 of the vessel, or can be held in the desired position in the collecting vessel 1 by means of static friction. It is further possible that, as indicated by the broken line in FIG. 2, the collecting vessel 1 on the inner wall 5 of the vessel has a web 22 protruding into the interior 3 of the vessel, which web can be formed all around the circumference or even only partially over extends the circumference of the collecting vessel 1. The scope is to be understood as the scope of the inner wall 5 of the vessel perpendicular to the central axis 7 of the vessel jacket. The latter embodiment offers the advantage that the carrier 21 can be removed from the collecting vessel 1 in a simple manner and is thus available for further processing for analysis of the respective substance to be detected or after removal of this substance for reuse.

The carrier 21 can be designed, for example, in the form of a preferably inert filter. It is thus possible to make the support 21 made of glass, e.g. To manufacture glass fibers, silicate materials, cellulose, latex etc.

3 shows another embodiment variant for the arrangement of the carrier 21 in the collecting vessel 1. The vessel jacket 6 has a dimensional change 23 in the direction of the vessel jacket center axis 7, which is designed such that the diameter of the collection vessel 1 decreases in the direction of the vessel bottom 10. The resulting constriction is designed in one step according to the example of FIG. 3, but it is also possible that this dimensional change 23 takes place continuously and thus the diameter of the vessel opening 4 decreases continuously in the direction of the vessel bottom 10. With the help of this change in dimension 23, it is possible to inject the carrier 21, which has a predeterminable diameter, at a specific height 24, measured from the vessel bottom 10

To hold the direction of the vessel opening 4 and therefore no further measures for holding the carrier 21 in the collecting vessel 1 are necessary. It can thus be achieved that the carrier 21 only rests on the dimensional change 23, and it is thus possible to simply remove the carrier 21 from the collecting vessel 1, for example with the aid of a tweezers.

As further shown in Fig. 3 by the dashed or dash-dotted line within the carrier 21, it is possible to divide the carrier 21 zonar and in these zones reagent or reagent gemi see 20 differently, so that a multi-component analysis as a result can be done. - lo -

It is of course also possible in this way to stack a plurality of carriers 21 on top of one another, so that a separation of the individual carriers is possible in a simple manner and the substances to be detected can subsequently be separated without the need for further separation steps.

If necessary, the substance to be detected, for example DNA, RNA, can of course be eluted from the carrier with the aid of an appropriate solvent, and it is subsequently also possible to reuse it by regeneration of the carrier.

The carrier 21 can have capillaries in its interior, for example in the manner of a filter, so that the liquid 2 to be examined is forced to flow through these capillaries and contact with the reagent or reagent mixture 20 can take place. These capillaries can furthermore be designed such that they have a diameter which enables at least partial separation according to the molecular weight of the constituents of the liquid 2. In this way it can be achieved, since the DNA or RNA molecules usually have a higher molecular weight than the remaining substances in the blood, that these molecules cannot enter the interior of the capillaries and thus a separation of the DNA or RNA molecules from the remaining constituents of the liquid 2 to be analyzed and thus fractionation by molecular weight is possible.

If the carrier 21 is of multilayer construction, in addition to the possibility that several individual components are analyzed in parallel, there is also the possibility that one layer for the lysis, another layer for the stabilization and a third layer for the qualitative or quantitative separation of the analyzing substance is used.

Additional layers for additional analysis steps are of course possible.

The carrier 21 according to FIGS. 2 and 3 can have the shape of a tablet, for example.

FIG. 4 shows an embodiment variant of the collecting vessel 1, the inside of the vessel 3 being the

Carrier 21 is arranged in the form of particles 25. These particles 25 can have permanent magnetic properties and can consist, for example, of an Fe ₂ O ₃ , Ni core or the like with a plastic or glass coating, and can be coated with the reagent or reagent mixture 20 over at least part of their outer surface. The shape of the particles 25 can be chosen as desired, for example spherical, lenticular, rod-shaped, etc. A suitable formation of the reagent or reagent mixture 20 on these particles 25 can result in an affinity separation of the substances present in the liquid 2, ie the reagent or reagent mixture 20 can be selected so that there is preferably a particularly high affinity for the substance to be analyzed , In order to bind the DNA or RNA molecules to these carriers 25, for example, it is possible to present a corresponding buffer in the interior 3 of the vessel, with the aid of which the blood sample can be stabilized, for example, and the cells can be lysed.

As already mentioned, it is possible, for example, that the reagent or reagent mixture 20 effects the lysing of biological cells and thus e.g. can stabilize nucleic acids (DNA, RNA). In this case the reagent or mixture 20 may contain a guanidinium salt, e.g. a guanidinium halide such as e.g. Guanidinium chloride, guanidinium thiocyanate, guanidinium carbonate, guanidinium nitrate, guanidinium acetate, guanidinium sulfate, guanidnium stearate, guanidinium dihydrogen or hydrogen phosphate or the like., Guanidinoalkylsulfate ester, chaotropic or rodium chloride, perchlorate, peroxide, potassium iodide, potassium iodide, potassium iodide, potassium iodide, potassium iodide, potassium iodide, peroxide / Phenol mixtures or the like. Contain. In addition, it is of course possible, in order to maintain certain environmental conditions, that this reagent or reagent mixture 20 furthermore contains buffer substances and / or detergents, such as e.g. Tris (2,3-dibromopropyl) phosphate, tris (hydroxymethyl) aminomethane, ethoxylates of 4- (l, l, 3,3-tetramethylbutyl) phenol, benzyltrimethylammonium hydroxide, (4- (2-hydroxyethyl) piperazino) -ethanesulfonic acid, 3-Moφholino-l-propanesulfonic acid, sodium dodecyl sulfate, polyoxyethylene derivatives of sorbitan esters, such as contains laurate, palmitate, stearate, tristearate or oleate. In addition, reducing agents such as e.g. ß-mercaptoethanol may be included.

It is also possible that the inner wall 5 of the vessel is coated with reagents in the solid state, e.g. with anticoagulants such as EDTA, the latter can also be presented as a solution.

Furthermore, the reagent or reagent mixture can contain 20 citrate and / or phosphate buffers, polydocanol as a detergent and dithiothreitol, TCEP as a reducing agent.

The guanidinium component can be present in a concentration between 0.05 M to 10 M and the buffer can be contained in the range between 1 mM and 500 mM. The detergent and the reducing agent can in turn be used in concentrations of 1 to 45% by weight or 0.1 to 15 wt .-% are present.

The pH is preferably in the acidic range, for example between 3 and 6.5, but can also assume values up to 8.

The advantage that can be achieved by the loosely placed particles 25 is a simple separation of the latter from the liquid matrix remaining in the collecting vessel 1. This separation can e.g. in such a way that, as indicated by the broken line in FIG. 4, a separation device 26 is introduced into the interior 3 of the vessel. In order to avoid an unwanted spilling of the liquid 2, it is possible to provide this separation device 26 with the closure device 11, it being possible to dispense with the septum 12. In particular, it is possible to design the closure device 11 as a cap in which the separation device 26 is held.

If particles 25 with permanent magnetic properties are selected as carriers 21, it is possible to bind these particles 25, provided the separation device 26 also has magnetic properties, to the separation device 26 due to the laws of magnetism, which is shown in FIG. 4 by the one magnetic field 27 indicated arrows 28 is shown schematically. The separation device 26 can either have permanent magnetic properties or it is also possible to measure the forces of the

To use electromagnetism.

After the particles 25 have been attached to the surface of the separation device 26, they can be removed from the liquid 2 and, for example, placed in a further collecting vessel 1, where the particles 25 are subsequently detached from the separation device 26 and e.g. the analysis is made possible by detaching the reagent or reagent mixture 20 with the substance from the carrier 21 and by subsequent detection of the substance.

Of course, it is also possible that the reagent or reagent mixture 20 remains on the particles 25 and only the substance to be analyzed, e.g. DNA, RNA, is eluted.

Of course, it is also possible for this separation device 26 to be designed differently, for example for it to be one on the side facing the vessel bottom 10 Device 29, as shown in Fig. 5, which has, for example, the shape of a network. The particles 25 can be held on this device 29, so that on the one hand the particles 25 remain on the device 29 when the separation device 26 is pulled out of the collecting vessel 1 and on the other hand it becomes possible for excess liquid 2 to remain in the interior 3 of the collecting vessel 1 through this network ,

The particles 25 can be a loose bed of possibly multi-layer individual particles, e.g. Glass balls, silica gel particles or the like., Be arranged in the collecting vessel 1. It is possible to arrange this loose bed at a predefinable height 24 from the vessel bottom 10, for which purpose a corresponding support device should be present in the collecting vessel 1. This support device can be designed, for example, as a perforated plate, which is held at a predefinable height 24 by suitable means, for example by arranging the web 22, but also by designing the collecting vessel according to FIG. 3.

Furthermore, it is possible that the particles 25 e.g. are encapsulated in their own envelope, which enables better cohesion of these particles 25 and should this envelope be suitable for the passage of liquid, i.e. for example, have a corresponding perforation. It is also possible that this cover consists of absorbent materials, for example cellulose fibers or the like. This encapsulation of the particles 25 in turn enables the latter to be removed easily, so that in a subsequent work process the substance or reaction products of the reagent or reagent mixture 20 to be analyzed can be separated, for example with the aid of an elution process, and is again the detection of the substance to be analyzed is possible without additional work steps.

6 and 7 show possible design variants for the carrier 21. This can e.g. comprise a carrier body surface 31, which may have the shape of a rod, a chain or the like, for example, and it is possible to coat this carrier body surface 31 with the reagent or reagent mixture 20. Corresponding precautions can be taken for enlarging the surface, for example by arranging surface enlargements 32 of at least approximately hemispherical shape. Of course, these surface enlargements 32 can have any shape adapted to the respective intended use.

These surface enlargements 32 can either be moved with the carrier body 30. be firmly connected or form part of the carrier body 30, for example by direct molding during the manufacturing process.

By means of these surface enlargements 32, it is also possible to zonarily divide the carrier body 30, whereby the arrangement of different reagent or reagent mixtures 20 on the carrier body 30 and thus a multi-component analysis is possible.

Furthermore, it is possible that the particles 25 are fastened in a suitable manner on the carrier body surface 31 and can in turn take them up separately in their own carrier body article, e.g. in the aforementioned case, which makes it easy

Removal of the particles 25 from the carrier body 30 becomes possible. The Trägerköφeφartikelaufnahme is preferably designed for the passage of the liquid 2, whereby the reaction with the reagent or reagent mixture 20 is possible.

It is also possible that these particles have 25 capillaries, on the one hand, for the

Reaction with the substance available surface of the particles 25 to enlarge or on the other hand a selective separation by molecular weight, i.e. according to molecular size.

Of course, the carrier bodies 30 shown in FIGS. 6 and 7 can have any suitable, arbitrary shape.

It proves to be advantageous if the carrier body 30 is elongated with two carrier body ends 33 arranged opposite one another. This makes it possible for the closure device 11 to be arranged on at least one carrier body end 33, any suitable connection technology between the carrier body 30 and the closure device 11 being able to be selected, or it is of course possible for the closure device 11 to be made in one piece with the carrier body 30 is. It can thus be achieved that the Trägerköφer 30 is inserted into the collecting vessel 1 so that an unintentional spilling of the contained therein by sealing the vessel opening 4 of the collecting vessel 1

Liquid 2 is prevented. Thus, for example, an intimate mixing of the liquid, for example by shaking, is possible without the person operating the collecting vessel 1 running the risk of possibly coming into contact with viruses and bacteria present in the liquid 2, for example HIV viruses, tuberculosis viruses etc. , For this purpose, the closure device 11 can be provided with a thread, for example, and can be used, for example, as a screw cap. be led. Likewise, it is of course possible to use appropriate frictional forces between the closure device 11 and the surface of the collecting vessel 1 and thus to design the closure device as a simple plug-in closure.

To improve the sealing of the vessel opening 4 by the closure device 1 1, it is of course possible that corresponding sealing elements 34 are arranged in the latter. These sealing elements 34 can be made from materials known from the prior art and, for example, consist of elastomers. Furthermore, it is possible that the closure device 11 additionally includes the septum 12. This enables the loaded carrier 21 to be removed from a first collecting vessel 1 and placed in a second collecting vessel 1 (not shown) and subsequent detachment of the substance to be examined by introducing an appropriate device, for example a cannula 19 (not shown). , through which septum 12 a suitable solvent can be brought into the collecting vessel 1 and there is in turn the possibility, due to the sealing device 11 sealing the collecting vessel 1, for better and faster detachment of the reagent loaded with the substance to be analyzed or Reagent mixture 20 from the carrier body 30 (or the species resulting from the reaction of the substance with the reagent or reagent mixture 20) to be used. In order to achieve a better distribution of a solvent to be supplied through the septum 12, the carrier body 30 can be provided with a channel 35 which extends from the septum 12 to at least approximately to the carrier body end 33 arranged opposite it (both the septum 12 and the channel 35 are indicated by dashed lines in Fig. 6).

Furthermore, it is possible that 35 distribution channels branch off from this channel, which extend as far as the carrier body surface 31, or else into the particles 12. It was on this

Reference mentioned that due to the affinity of RNA or DNA molecules for glass, it is of course possible to use carrier body 30 or carrier 21 which are not coated with the reagent or reagent mixture 20.

Carrier bodies 30 made of a silicate material, cellulose, plastic or the like can also be used.

There is also the possibility of producing the carrier body 30 from a gel, for example an agarose sail. Furthermore, it is possible that a corresponding reagent or reagent mixture 20 is placed in the collecting vessel 1, in which the carrier body 30 is inserted, with the aid of which an appropriate preparation of the liquid 2 can take place, for example the stabilization of the liquid 2, for example blood , and / or the lysis of the cells.

The carrier 21 or carrier body 30 can, however, also be made from an open-pore or closed-cell foam which is impregnated with a corresponding reagent or reagent mixture 20 and which allows the liquid 2 to be analyzed to pass through. This foam can, for example, be made soft and it is possible to manufacture it from polyurethane. Of course, semi-hard to hard are also

Possible designs of the foam.

8 partially shows a collecting vessel 1, in which the reagent or reagent mixture 20 is not presented as a solid, but in the form of a corresponding liquid 36 or a liquid mixture and / or a solution of solids required for analysis in the corresponding solvents. The advantage that can be achieved with this is that by supplying or introducing the liquid 2 to be analyzed into the collecting vessel 1, the reagent or reagent mixture 20 does not first have to be dissolved by the liquid 2 in order to react with the analyzing substance to be available. Since it is also possible to measure the vacuum prevailing in the collecting vessel 1 in such a way that a predeterminable amount of liquid 2 is sucked into the collecting vessel 1, the concentration of the reagent or reagent mixture 20 can always be in excess, so that the reaction is rapid analyzing substance or substances can take place with the reagent or reagent mixture 20 and, as a result, stabilization of DNA or RNA molecules, for example, may not be necessary. This information regarding the concentration can of course also apply in the event that the reagent or reagent mixture 20 is in the solid state of matter.

Of course, it is necessary for the closure device to be designed accordingly to maintain the vacuum in the collecting vessel 1 over a longer period of time, as is necessary, for example, for storing this collecting vessel 1.

9 again shows an embodiment variant of the collecting vessel 1 according to the invention, in which the carrier 21 is arranged in a collecting vessel extension 37, this collecting vessel extension 37 preferably opening collecting vessel extensions 38. has. One of these collecting vessel extension openings 38 is connected to the vessel jacket end faces 8 opposite the vessel bottom 10 and can for example be connected to the vessel jacket 6 by means of a thread or other connection methods can of course be selected, for example this connection can be designed as a plug connection. In any case, it should be ensured that this connection is so tight that the vacuum prevailing in the collecting vessel 1 is not broken down by air entering.

The carrier 21 can now either be connected to this collecting vessel extension 37 in a fixed manner, or there is the possibility of mounting this carrier 21 on at least part of the

To allow vessel jacket end faces 8 to rest and only to prevent the carrier 21 from sliding sideways by means of the collecting vessel extension 37.

The second collecting vessel extension opening 38 is in turn preferably closed with a closing device 11 - expediently with the septum 12. This makes it possible that, by perforating this septum 12, for example with a cannula 19, not shown, the liquid 2 to be analyzed initially encounters the carrier 21, which in turn can contain the reagent or reagent mixture 20, in the carrier after it has entered the collecting vessel 1 21 the required reactions take place and finally the liquid matrix that is not required runs into the lower part of the collecting vessel 1. For further

Analysis or to detach the reaction products or the substance to be analyzed from the carrier 21, this can either, provided that the carrier 21 is connected to the collecting vessel extension 37 in a motionless manner, be placed together with the latter on a further collecting vessel 1 and, for example, again through the septum 12 Closure device 11 can be acted upon with an appropriate solvent, so that the one to be examined

Substance or the substance mixture to be examined runs into the further collecting vessel 1 and is available there for the further process steps or the detection.

If, however, the carrier 21 is not connected to the collecting vessel extension 37 in a fixed manner, it is possible to transfer the carrier 21 separately into a further collecting vessel 1 or to feed it directly to a final detection of the substances to be determined.

In this way it is possible to assemble a complete analysis kit 39 for the analysis of liquids of biological origin or biological matrices, such as this is shown for example in Fig. 10. Several collecting vessel extensions 37 are arranged on the collecting vessel 1 in the manner already described, these collecting vessel extensions 37 again containing the carrier 21, as already described. A closure forms the closure device 11 already described, which on the one hand maintains the vacuum built up in the collecting vessel 1 and on the other hand enables the liquid 2 (not shown) to be supplied. In this way, by arranging several carriers 21 one above the other, practically simultaneously, several substances or substance mixtures to be analyzed can be separated from the liquid 2 and are thus available for further analysis.

However, this arrangement according to FIG. 10 can also be used to separate the reagent or reagent mixture 20 (not shown) into individual stages, so that the individual process steps required, such as stabilization and / or lysis of the cell and / or the molecules to be examined are immobilized on different levels.

Of course, it is also possible in this case that the carrier 21, as already described, has a multi-layer design.

Another embodiment variant for such an analysis kit 39 is shown in FIG. 11. The

In this case, the collecting vessel 1 again consists of the vessel jacket 6, although a further vessel opening 4 is present instead of the previously described vessel bottom 10.

However, it is also possible, as shown in dashed lines in the lower part of FIG. 11, that a further vessel opening 4, e.g. with the cannula 19 is arranged.

The two vessel openings 4 are preferably closed by the closure device 11, which contains the septum 12. On the one hand, it can thereby be achieved that the liquid 2 to be analyzed is supplied to the interior 3 of the vessel via a corresponding device, for example the cannula 19. On the other hand, however, it is also possible to connect several of these collecting vessels 1 to one another via a cannula 19, as shown in FIG. 11.

Such a design makes it possible to arrange different, not shown, reagents or reagent mixtures 20 in the different collecting vessels 1, where when this can be done either in solid or liquid form, with the aid of which a separation according to constituents or decomposition products of liquid 2 can take place. After separation of the individual collecting vessels 1, they are thus automatically available for further analysis without additional manipulation steps.

Of course, it is also possible with this variant of the analysis kit 39 to arrange carriers 21 in the collecting vessels 1, which can be removed through the respective vessel opening 4, but are not shown in FIG. 11.

With this method it is also possible to equip the analysis kit 39 in such a way that one of the

Collection vessels 1, the elution agent required for the elution of the substance to be analyzed is presented from the carrier 21 and, after the latter collection vessel 1 has been joined together with one or more collection vessels 1 containing the carriers 21, it runs over the respective carrier 21 and thus causes the elution.

It is also possible that the analysis kit 39 is designed so that several of these collection vessels contain saline solutions, e.g. Containing cesium chloride solutions, of different densities, which enables the liquid 2 to be analyzed to be separated by centrifugation, preferably at high g values, using a cesium chloride gradient. It goes without saying that the corresponding collecting vessel 1 with the desired cesium chloride concentration is connected to the collecting vessel of the previous cesium chloride concentration after each centrifugation step.

FIG. 12 again shows a variant of the collection vessel 1, which can be provided with 2 vessel openings 4, which have the closure device containing the septum 12

1 1 are closed. A device 40 is arranged in the interior 3 of the vessel, with the aid of which a physical separation of the constituents of the liquid 2 can be effected during or after the centrifugation. The density of this device 40 is such that the density of one of the two phases formed during centrifugation is greater and the density of the other phase is lower, so that this device 40 on the

The interface of the two phases comes to lie.

In order to enable phase separation, this device 40 can have a schematically indicated device opening 41 which closes again after centrifugation and by means of which the phase with the lower specific weight is made possible in to climb the upper part of the collecting vessel 1.

However, the device 40 can also be designed such that it does not abut the entire circumference of the vessel jacket 6 during centrifugation, i.e. on the inner wall 5 of the vessel, so that the phase with the lower specific weight between the inner wall of the vessel

5 and the device 40 can get into the upper part of the collecting vessel 1.

It goes without saying that the tube-like connections shown in FIG. 12, for example the cannulas 19, via which a connection of several collecting vessels 1 can take place, are not attached to the collecting vessel 1 or in the septum 12 during centrifugation.

Due to the separation of the two phases and the accessibility of the collecting vessel 1 via at least two vessel openings 4, the two phases can be separated from one another in a simple manner, so that aftertreatment of at least one of the phases with a corresponding reagent or reagent mixture 20 in the same or with one this collecting vessel 1 connected further collecting vessel 1 is possible.

However, the two vessel openings 4 can also be used to access a carrier 21 arranged in the collecting vessel 1 but not shown in FIG. 12 from both sides.

If the phase separation leads to more than two different phases, it is possible, as already described, to arrange at least one further vessel opening 4 along the vessel jacket 6, which in turn can be closed with a corresponding closure device 11.

In the event that several collecting vessels 1, as shown in Fig. 10, are connected to an analysis kit 39, there is the possibility that the vacuum in the collecting vessel 1, into which the liquid 2 to be analyzed last enters, is selected so that the liquid 2 is sucked through all of the present receptacles 1.

13 shows an embodiment variant of the collecting vessel 1, in which a conventional collecting vessel, for example a blood collection tube, is connected to a collecting vessel 1 according to the invention. The connection can again be made via a thread, a plug closure or the like, or it is also possible for a plurality of collecting vessel extensions 37 (not shown in FIG. 13) to be arranged between the two collecting vessels are.

This configuration makes it possible to collect the liquid 2 to be analyzed first in the conventional collecting vessel 1 and to stabilize it there for any later analysis and to store it there.

In turn, the carrier 21 can be arranged in the collecting vessel 1 according to the invention and it becomes possible that the liquid 2 passes through the carrier by a tilting movement of this arrangement and the substance to be analyzed thus adheres to the carrier and is available for further analysis.

Of course, the reverse process is also possible, namely that the reagent or reagent mixture 20 arranged on the carrier 21 selectively binds or reacts with those substances which are not to be analyzed, so that only the substance mixture to be analyzed or the substance to be analyzed by passes through the carrier 21.

Fig. 14 shows an arrangement of the carrier 21 in the course of the previously described tubular connection, for example the cannula 19, with the aid of which e.g. a patient's blood can be drawn. This carrier 21 can in turn be loaded with the reagent or reagent mixture 20.

It is possible in this way, since the carrier is now not connected to the collecting vessel 1 or is contained in the interior 3 of the vessel, the substance to be analyzed simply flows from the liquid matrix, which after passing through the carrier 21 into the collecting vessel 1, with which the cannula 19 is connected via the septum 12, or it is therefore not necessary to open the collecting vessel 1 in order to remove the carrier 21 from the interior 3 of the vessel and in turn a higher level of safety for the operating personnel can thus be achieved.

FIG. 15 shows an embodiment variant of the collecting vessel 1, in which the carrier 21 is connected directly to the septum 12 of the closure device 11, for example is molded onto it. Of course, other connection methods are possible.

In order to enable the reaction of the substances contained in the liquid 2 with the reagent or reagent mixture 20 contained on the carrier 21, the tube-like one should Connection, for example the cannula 19, cannot be pushed entirely through the carrier 21.

With this embodiment variant, it is also possible to remove the carrier 21 with the substances to be analyzed by removing the closure device 11 from the collecting vessel 1.

16 finally shows an embodiment variant in which an inner collecting vessel 1 is contained in an outer collecting vessel 1 or can be inserted into the outer collecting vessel 1.

The inner collecting vessel 1 has at least one vessel jacket opening 42 in the vessel jacket 6. The connection of the interior to the outer collecting vessel 1 can take place, for example, by means of a slide closure 43, which is arranged at the vessel opening 4 of the outer collecting vessel 1 or is screwed to the vessel jacket 6 of the outer collecting vessel 1, and this sliding closure 43 can, for example, be made of known Quickfit be carried out. The inner collecting vessel 1 can be pushed into the outer collecting vessel 1 through the central opening which this slide closure 43 has.

If a liquid 2 to be analyzed is placed in the outer collecting vessel 1, the insertion of the inner collecting vessel 1 into the outer collecting vessel 1 generates a corresponding pressure on the liquid 2, so that this, if a diameter 44 is chosen smaller than a collecting vessel inner diameter 45 of the outer collecting vessel 1, whereby the wall thickness of the inner receptacle 1 must of course also be taken into account, escapes into the gap between these two receptacles 1 and subsequently enters the interior 3 of the inner receptacle 1 through the vessel jacket openings 42 in the inner receptacle 1. If the inner receptacle 1 is now coated on the outside with a reagent or reagent mixture 20, the liquid 2 or its constituents can be on the way from the outer receptacle 1 into the inner receptacle 1

Reagent or reagent mixture 20 react so that individual components or cleavage products or mixtures of substances can be deposited thereon.

If a slide seal 46 is now designed such that when the inner one is pulled out of the outer collecting vessel 1, the reagent or reagent mixture 20 with the one on it located substance or the mixture of substances is stripped from the outer surface of the inner container 1, so an automatic separation of the substances to be analyzed can take place. Of course, if the reagent or reagent mixture 20 should interfere with the analysis, especially if it is selected as the carrier 21, it must be separated from the substances to be analyzed.

This embodiment variant of the collecting vessel 1 or analysis kit 39 is also particularly suitable if the liquid to be analyzed is temporarily stored and the analysis is only carried out at a later point in time.

The reagent or reagent mixture 20 can be selected or composed such that the lysis and / or stabilization of individual components of the liquid 2 or the biological matrix is effected. It is of course also possible that this reagent or reagent mixture 20 or part of this reagent mixture 20 is an extracting agent, e.g. a phenolic solution, a phenol / chloroform mixture or the like, in order to use it, for example, to separate proteins from the DNA, with the aid of which part of the liquid 2 or the biological matrix is converted into this extractant, as is customary with extractants. In accordance with Neernst's law, the extractant should be selected so that the concentration of the substance to be analyzed in the remaining liquid matrix is almost close to zero by a one-step "shaking process".

For stabilization, especially when the liquid 2 is blood, the reagent or reagent mixture 20 can contain a guanidinium salt.

Furthermore, the reagent or reagent mixture 20 can at least partially contain cesium chloride in order to be able to carry out a cesium chloride centrifugation known from the prior art.

The reagent or reagent mixture 20 can also be selected so that the liquid 2 or components of the biological matrix are divided into several phases.

It is also possible for the reagent or reagent mixture 20 to selectively precipitate individual constituents of the liquid 2 or of the biological matrix, in order, for example, to thereby purify the nucleic acids. Thus, the reagent or reagent mixture 20 can be selected such that the DNA or RNA molecules or cleavage products thereof be put down. On the other hand, however, it is also possible that, for example, the so-called salting-out effect or changes in the pH value lead to a selective precipitation of proteins.

For example, it is possible for one of the collecting vessels 1 of the invention to contain isopropanol or ethanol in order to additionally concentrate the DNA or RNA molecules or the cleavage products thereof or individual other constituents of the liquid 2 or of the biological matrix. If it should be expected that the concentration of the nucleic acid is low, it is also possible to mix an inert carrier 21 with the reagent or reagent mixture 20, for example

Glycogen to increase the efficiency of precipitation formation.

The reagent or reagent mixture 20 can also consist of lithium chloride or contain lithium chloride. Lithium chloride is e.g. preferably used when only RNA is to be precipitated and not proteins or DNA.

In the case of the permanent magnetic carrier 21, it is possible that these are coated, for example, with streptavidin, for example for the analysis of poly (A) ⁺ mRNA. Other compositions of the reagent or reagent mixture 20 are possible in order to specifically deposit other nucleic acids or to bind them adsorbently. However, it is also possible for the reagent or reagent mixture 20 to be a gel, for example an agarose gel, a plastic gel or the like, so that gel filtration can subsequently be carried out, the gel representing a matrix with a defined pore size. Such molecules, whose diameter is smaller than the defined pore size, can therefore diffuse into these pores, whereas larger molecules are prevented from entering and these can thus be separated off.

The molecules that have penetrated into the pores can subsequently be delayed according to their molecular size, e.g. be eluted according to increasing molecular size.

In addition to the above-mentioned selection for the reagent or reagent mixture 20, the latter can of course be chosen so that the nucleic acids can be purified, for example by chromatography, electrophoresis, centrifugation, affinity separation or the like. As already mentioned, a support 21 made of glass is used, since the DNA molecules have an affinity for glass and can thus be separated from proteins and RNA, for example. There is also the possibility of forming the reagent or reagent mixture 20 from endo- and / or exonucleases in order to enable the structure of the DNA or RNA to be clarified. The endonucleases can be, for example, deoxy and / or ribonucleases, both enzyme groups cleaving DNA or RNA, but only if they consist of 3 ', 5' phosphorus diester bonds. For example, the two DNAses I or II can be used to convert high molecular weight DNA into nucleotides, with DNAse I leading to tri- and tetradesoxinucleotides with a 5'-phosphate group. DNAase II forms oligode soxinucleotides with 3 'phosphate ends.

Of course, it is also possible for the reagent or reagent mixture 20 to form an appropriate buffer in order to provide a medium suitable for analysis or deposition, stabilization, immobilization, etc.

Furthermore, it is possible to carry out an indirect detection with the aid of the reagent or reagent mixture 20, for example with the biotin / streptavidin amplification method, this reagent or reagent mixture should contain at least 2 antibodies.

The use of the collecting vessel 1 or the carrier 21 or the analysis kit 39 can now be done in such a way that the reagent or reagent mixture 20 already during the introduction of the liquid 2 into the collecting vessel 1 this or the biological matrix or at least some Separates components from it and / or immobilizes and / or stabilizes. For further separation of the liquid 2, several collecting vessels 1 can be connected to one another, as has already been explained. If the reagent or reagent mixture 20 does not already cause separation of individual constituents from the liquid 2 or the biological matrix, it is possible with the aid of the carrier body 30 to effect this separation by introducing it subsequently into the collecting vessel.

17 shows a further exemplary embodiment of the collecting vessel 1 according to the invention. Although it has a single layer, in this and all the variants described above there is the possibility of carrying out the collecting vessel 1 in one or more layers, ie that the vessel jacket 6 as a laminate of different plastics can be executed. It is also possible to select the plastics in such a way that at least one takes over the supporting function of the collecting vessel 1 and a further plastic is designed to be correspondingly tight with respect to the permeation of liquids and / or gases. For example, it is possible that the part of such a laminate made of polypropylene facing the interior 3 of the vessel len and the outer layer of this laminate, which is preferably molded onto it, consists of polyethylene therapy phthalate.

There is also the possibility of not designing the vessel jacket 6 of the collecting vessel 1 as a laminate of different plastics, but in the form of at least two individual vessels pushed into one another, which in turn are made of different plastics, e.g. meet the above requirements.

In the embodiment variant according to FIG. 17, it is shown how, in a simple manner, the particles 25, which in turn can be designed as permanent magnetic particles with or without a coating, together with the substance to be analyzed from the matrix, e.g. the blood drawn. For this purpose, the closure device 11, for example a screw cap, can have a finger-like extension 47, preferably made of plastic, glass or the like. This extension 47 is preferably made of transparent material and has an opening 48 in the direction of the surroundings. The side of the extension 47, which is opposite this opening 48 and preferably adjacent to the bottom 10 of the vessel jacket, is preferably closed.

With the help of this extension 47, it becomes possible to insert a rod 49 into it, for example, which can have magnetic properties. Under the influence of the magnetic field emanating from this magnetic rod 49, the particles 25 are now bound to an outer surface 50 of the extension 47 and this makes it possible to remove these particles 25 together with the substance to be analyzed from the liquid 2, for example by the closure device 1 1 is removed.

In a further embodiment, it is possible for a holding device 51 to be arranged on the closure device 11, with the aid of which the rod 49 can be arranged in the extension 47 at least approximately immovably. This holding device 51 can be designed, for example, in the manner of a seal, with the frictional forces between the

Holding device, for example an elastomer arranged in this, and the rod 49 a corresponding holder can be achieved. The holding device 51 is indicated by dashed lines in FIG. 17 and, for example, the locking device 11 can in this case again be designed as a quick fit. Of course, it is also possible in this embodiment variant of the collecting vessel 1 that the closure device 11 comprises the septum 12 (not shown in FIG. 17), so that subsequent replacement of the closure device by the type described in FIG. 17 is not necessary and can be done, for example the septum 12 may be formed in a ring around the extension 47.

In the event that this collection vessel 1 according to FIG. 17 is used for taking blood with simultaneous lysis of the cells and stabilization of the nucleic acids in order to prevent the latter from being broken down, a solution, as already described, can preferably be used in the collection vessel 1. from a chaotropic substance, for example a guaninium salt, a buffer, a detergent and preferably a reducing agent. In this case, it may be sufficient if the particles 25 are only designed as permanent magnets and the latter is not coated, provided that a selective analysis for certain nucleic acids is not necessary. For this purpose, as already mentioned, these particles 25 can have a multi-layer design and can comprise, for example, a magnetic core made of Fe ₂ O ₃ or nickel or the like, which is surrounded by a further layer, for example made of plastic or glass. On the other hand, it is of course possible for the layer structure to be chosen differently, namely from a carrier substance for the core, which is coated with a magnetic material and in turn a polymer layer is arranged above it.

Instead of a closure device 11, as shown in FIG. 17, with the extension 47, it is possible to use a conventional closure device 11, provided with or without a septum 12, and the separation of the preferably magnetic particles 25 with the aid of an externally attached to the Make collecting vessel 1 listed magnets. In this case the

Removal of the liquid residual substance by removing the closure device 1 1 and emptying the collecting vessel 1.

It is also possible to design the collecting vessel 1, in particular the closure device 11, in such a way that the extension 47 is not formed on the closure device 11, but rather can be removed. This makes it possible to automatically remove this extension 47 together with the magnets and the particles 25 adhering to the extension 47, for example in an automatic system for analysis, and for example to remove the particles 25 with the substance to be analyzed adhering to them Transfer the reaction vessel. The particles 25 can be detached from the extension 47, for example by pulling the rod 49 out of the extension 47. In order to prevent the particles 25 from being wiped off when the extension 47 is removed from the closure device 11 in this embodiment variant, it is possible to remove a portion 52 of the closure device 11 around the extension 47 together with the latter. This partial area 52 is indicated by dash-dotted lines in FIG. 17, and it is possible, for example, to provide a predetermined breaking point in the closure device 11.

In addition to the materials for these particles 25 already mentioned, the latter can also consist of porous glass, for example of silicon dioxide. This has the advantage that the chemical inertness of this material means that organic and aqueous solvents can be used over a wide pH range even in the presence of aggressive reagents or reagent mixtures 20. Furthermore, both these and all the other particles 25 described can be coated. For example, in the case of the glass particles, the surface can be provided with hydrophilic and / or hydrophobic reactive groups which e.g. are bound to the porous glass via covalent siloxane bonds.

There is also the possibility of producing these particles 25 from polystyrene.

The diameter of the particles 25 can be in the range between 0.1 μm and 250 μm, for example between 0.4 and 100 μm, in particular between 1.0 μm and 50 μm. For individual applications, the largest grain can also have a diameter of over 500 μm.

In a further embodiment variant, it is possible to equip the closure device 11 with a permanent magnet or, for example, an electromagnet. This makes it possible to bind the permanent magnetic particles 25 to the closure device 11 by simply turning the collecting vessel 1 around, and these can be safely removed from the collecting vessel together with the closure device 11. The use of an electromagnet is particularly advantageous, for example by installing a coil in the closure device 11 with corresponding connections for energy supply, since this can prevent the particles 25 from inadvertently adhering to the closure device 11 before they are actually used. By appropriate design of the closure device 11, subsequent cleaning of the latter and possibly sterilization is possible and the closure device 11 can thus be used several times. 18 and 19, a further exemplary embodiment of the collecting vessel 1 according to the invention with a closure device 11 is shown schematically in simplified form. In this embodiment variant, the closure device 11 serves to hold at least two analysis rods 53, which can optionally be coated with a wide variety of reagents or reagent mixtures 20. This enables a multi-component analysis.

These analysis rods 53 can either be connected to the closure device 11 so that they cannot move, so that they can be removed from the liquid 2 together with the closure device 11 and the substances to be analyzed. On the other hand, it is of course possible for these analysis rods 53, as indicated by the broken lines in FIG. 18, to be held by holding devices 51.

As FIG. 19 shows better, these analysis rods can be arranged symmetrically around the preferably available septum 12 in the closure device 11, so that the various substances to be analyzed are already separated by this septum 12, in particular by a cannula 19 (in FIG 19 not shown) begins.

These analysis rods 53 are preferably designed as already described carriers for reagents or reagent mixtures 20 and this embodiment variant of the collecting vessel 1 according to the invention can be used, for example, for the qualitative analysis of e.g. Blood are used. It is possible for the reagents or reagent mixtures 20 to include a wide variety of, preferably color indicators, which means that the presence or absence of the substance sought can be inferred solely from the color change that has occurred or not.

It is also possible that in the event that the closure device 11 is designed as a magnetic carrier, pyroelectric or piezocrystals, for example, are arranged in it, so that as a result the current required for a corresponding electromagnet is generated by increasing the temperature or by pressurization.

For the sake of order, it should finally be pointed out that, for a better understanding of the construction of the collecting vessel, the carrier and the analysis kit, these or their components have been partially shown to scale and / or enlarged and / or reduced. The object on which the independent inventive solutions are based can be found in the description.

Above all, the individual in Fig.l; 2; 3; 4; 5; 6; 7; 8th; 9; 10; 1 1; 12; 13; 14; 15; 16; 17; 18, 19 shown the subject of independent, inventive

Form solutions. The relevant tasks and solutions according to the invention can be found in the detailed descriptions of these figures.

REFERENCE NUMBERS

Collecting vessel 41 device opening liquid 42 vessel jacket opening vessel interior 43 sliding closure vessel opening 44 diameter vessel inner wall 45 collecting vessel inner diameter vessel jacket 46 sliding closure seal vessel jacket central axis 47 extension vessel jacket end face 48 opening vessel jacket end face 49 rod vessel bottom 50 surface sealing device 51 holding device septum 52 partial area outer jacket 53 test tube wall tube outer tube change inner tube diameter changing tube Particle Separation Device Magnetic Field Arrow Device Carrier Body Carrier Body Surface Surface Enlargeranj Carrier Body Sealing Element Channel Liquid Collection Vessel Extension Collection Vessel Extension Opening Analysis Kit

Claims

Patent claims 1.Collecting vessel (1) for liquids (2), in particular containing biological origin or biological matrices, e.g. Blood collection tube, with one of a vessel jacket (6) and optionally a vessel bottom (10), which (r) an inner wall of the vessel (5) form (t), at least partially surrounded vessel interior (3), which is accessible through at least one vessel opening (4), characterized in that in the vessel interior (3) at least one reagent or reagent mixture (20) with which at least some Components of the liquid (2) or the biological matrix or fission products of these components can be separated and / or immobilized and / or stabilized, arranged on a carrier (21) or that the carrier (21) through the inner wall of the vessel (5) and / or a closure device (11), preferably provided with a septum (12), is formed for the vessel opening (4) or that the carrier (21) forms the reagent or reagent mixture (20). 2. Collection vessel according to claim 1, characterized in that the reagent or Reagent mixture (20) selective for nucleic acids, nucleic acid mixtures or nucleic acid components, e.g. Is mononucleotides, oligonucleotides. 3. Collecting vessel according to claim 1 or 2, characterized in that the reagent or reagent mixture (20) is selective for nucleic acid-degrading components of the liquid (2) or the biological matrix, e.g. Proteins, metabolites, nucleases, enzymes or the like. 4. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is the lysis of the cells of the Components of the liquids (2) of biological origin or the biological matrix. 5. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is in the solid state. 6. A collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is present as a liquid (36) or liquid mixture and / or solution. 7. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) causes the hybridization of individual components of the liquid (2) of biological origin or the biological matrix. 8. A collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) or part of the reagent mixture (20) is an extraction agent, e.g. a phenolic solution, a phenol / chloroform mixture. 9. A collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) has at least one guanidium salt, e.g. Guanidinium chloride, guanidinium thiocyanate, guanidinium carbonate, guanidinium nitrate, guanidinium acetate, guanidinium sulfate, guanidinium stearate, guanidinium dihydrogen phosphate, guanidinium hydrogen phosphate, guanidinoalkysulfate ester contains. 10. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) contains cesium chloride. 11. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) causes a separation of the liquid (2) of biological origin or the components of the biological matrix into several phases. 12. A collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is a precipitation of individual components of the liquid (2) of biological origin or the biological matrix, e.g. DNA, RNA molecules, cleavage products thereof. 13. A collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) contains ethanol and / or lithium chloride. 14. Collecting vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is concentrated the individual components of the liquid (2) of biological origin or the biological matrix, for example DNA, RNA molecules, cleavage products thereof. 15. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is a gel, e.g. is an agarose sail, a plastic gel. 16. Collection vessel according to one or more of the preceding claims, characterized in that the reagent or reagent mixture (20) is arranged at least in regions as a coating on the inner wall of the vessel (5). 17. A collecting vessel according to one or more of the preceding claims, characterized in that the carrier (21) is in the form of a preferably inert filter. 18. Collecting vessel according to one or more of the preceding claims, characterized in that the filter by a loose bed of individual particles, e.g. Glass balls, silica gel particles or the like. Is formed. 19. Collecting vessel according to one or more of the preceding claims, characterized in that the filter has capillaries. 20. Collecting vessel according to one or more of the preceding claims, characterized in that the capillaries have a diameter which enables an at least partial separation according to the molecular weight of the constituents of the liquids (2) of biological origin or the biological matrix. 21. A collecting vessel according to one or more of the preceding claims, characterized in that the carrier (21) made of glass, e.g. Glass fibers, silicate materials, cellulose, latex, plastic, foam. 22. A collecting vessel according to one or more of the preceding claims, characterized in that the carrier (21) is in the form of particles (25) with, for example, permanent magnetic properties. 23. A collecting vessel according to one or more of the preceding claims, characterized in that the carrier (21) or the reagent or reagent mixture (20) is accessible via two opposing closure devices (11). 24. Collecting vessel according to one or more of the preceding claims, characterized in that the carrier (21) or the reagent or reagent mixture (20) is accessible via at least one further vessel opening (4) in the vessel jacket (6) along the center axis of the vessel. 25. Collection vessel according to one or more of the preceding claims, characterized in that a device (40) is arranged in the interior of the vessel (3), which physically separates the components of the liquid (2) of biological origin or the biological matrix, e.g. after centrifugation. 26. Collecting vessel according to one or more of the preceding claims, characterized in that the vessel jacket (6) and / or the vessel bottom (10) is formed by a plastic with polar or polarizable groups. 27. Carrier (21) for reagents or reagent mixtures (20) with a carrier body (30) with a carrier body surface (31), in particular for a collecting vessel (1) according to one or more of claims 1 to 31, characterized in that on the Carrier body (30) a reagent or reagent mixture (20) is arranged with which at least individual components of the liquid (2) or a biological matrix or fission products of these components can be separated and / or immobilized and / or stabilized. 28. A carrier according to claim 27, characterized in that the carrier body surface (31) is coated with the reagent or reagent mixture (20). 29. A carrier according to claim 27 or 28, characterized in that the carrier body (30) consists of individual particles (25). 30. Carrier according to one or more of claims 27 to 29, characterized in that the particles (25) have capillaries. 31. Carrier according to one or more of claims 27 to 30, characterized in records that the Trägerköφer (30) is formed in the form of chain links. 32. Carrier according to one or more of claims 27 to 31, characterized in that the carrier body (30) is a holding device, e.g. has a rod to which the particles (25) are attached. 33. Carrier according to one or more of claims 27 to 32, characterized in that the particles (25) are designed as permanent magnets. 34. Carrier according to one or more of claims 27 to 33, characterized in that the particles (25) are held by a carrier body. 35. Carrier according to one or more of claims 27 to 34, characterized in that the Trägerköφeφartikelaufnahme is grid-shaped. 36. Carrier according to one or more of claims 27 to 35, characterized in that a mesh size of the Trägerköφeφartikelaufnahme is smaller than the diameter of the particles (25). 37. Carrier according to one or more of claims 27 to 36, characterized in that the carrier body (30) is elongated with two mutually opposite carrier body ends (33). 38. Carrier according to one or more of claims 27 to 37, characterized in that a closure device (11) is arranged on one of the carrier body ends (33). 39. Carrier according to one or more of claims 27 to 38, characterized in that the closure device (11) is designed so that the vessel opening (4) of a collecting vessel (1) can be closed, e.g. is designed as a screw cap or plug closure or is at least partially provided with a thread (16). 40. Carrier according to one or more of claims 27 to 39, characterized in that the closure device (11) comprises a septum (12). 41. Carrier according to one or more of claims 27 to 40, characterized in that the carrier body (30) is provided with a channel (35) which extends from the septum (12) to at least approximately to the carrier body end (33) arranged opposite it extends. 42. Carrier according to one or more of claims 27 to 41, characterized in that from the channel (35) branch channels which extend to the carrier body surface (31). 43. A carrier according to one or more of claims 27 to 42, characterized in that the carrier body (30) is a gel. 44. Carrier according to one or more of claims 27 to 43, characterized in that the carrier body (30) consists of glass, a silicate material, cellulose or a plastic, foam. 45. Analysis kit (39) for analyzing liquids (2) of biological origin or biological matrices containing at least one collecting vessel (1) for holding the liquid (2) or components of the liquid (2) and / or the biological matrices and / or at least one carrier (21) for receiving a reagent or reagent mixture (20), characterized in that the collecting vessel (1) according to one or more of claims 1 to 31 and / or the carrier (21) according to one or more of the claims 32 to 52 is / are formed. 46. Analysis kit according to 45, characterized in that at least one further collecting vessel via a tubular connection, e.g. Cannulas (19) are connected to the collecting vessel (1). 47. Analysis kit according to spoke 45 or 46, characterized in that a septum (12) is arranged at the connection of the tubular connection to the collecting vessel (1). 48. Analysis kit according to one or more of claims 45 to 47, characterized in that at least some of several collecting vessels (1) contain different reagents or reagent mixtures (20), with the aid of which a separation according to constituents or decomposition products of the liquid ( 2) of biological origin or biological Matrix is done. 49. Analysis kit according to one or more of claims 45 to 48, characterized in that a collecting vessel (1) an elution means for eluting the components or fission products of the liquid (2) of biological origin or the biological matrix bound to the carrier (21) contains. 50. Analysis kit according to one or more of claims 45 to 49, characterized in that the collecting vessels (1) contain a saline solution, e.g. a cesium chloride solution, the density of these solutions in the individual collecting vessels (1) being different. 51. Process for the separation and / or immobilization and / or stabilization of components or fission products of a liquid (2) of biological origin or containing biological matrices, e.g. Blood, the liquid (2) in a collecting vessel (1) is collected, characterized in that during the introduction of the liquid (2) the liquid (2) or the biological matrix or at least individual components thereof are separated and / or immobilized in the collecting vessel (1) with the aid of a reagent or reagent mixture (20) which is placed in the collecting vessel (1) and / or be stabilized. 52. The method according to claim 51, characterized in that the liquid (2) or the biological matrix or at least individual components is separated by the connection of at least two collecting vessels (1). 53. The method according to claim 51 or 52, characterized in that for the separation and / or immobilization and / or stabilization of constituents or fission products of a liquid (2) of biological origin or biological matrices containing a carrier (21) in the collecting vessel (1) , in particular according to one or more of claims 27 to 44. 54. The method according to one or more of claims 51 to 53, characterized in that the carrier (21) coated with the reagent or reagent mixture (20) contains biological substance or biological matrices after introduction of the liquid (2) the collecting vessel (1) is inserted.