CN113252915A

CN113252915A - Full-automatic immunoassay device

Info

Publication number: CN113252915A
Application number: CN202010088467.1A
Authority: CN
Inventors: 李颖; 吴勇
Original assignee: Zhongshan Taihui Biotechnology Co ltd
Current assignee: Shanghai Wuti Biotechnology Co.,Ltd.
Priority date: 2020-02-12
Filing date: 2020-02-12
Publication date: 2021-08-13

Abstract

The present disclosure relates to a fully automatic immunoassay device, which is configured to directly perform immunoassay detection on the whole blood sample without preprocessing the whole blood sample. The device includes a detection component, a sample supply component, a reagent supply component and a reaction cup supply component. The detection mechanism of the detection assembly includes an excitation light source and a photon detector. One or more reagent components and a target in the whole blood sample to be detected immunoreact in a cuvette at the detection assembly to produce a long-persistent luminescent complex. The excitation light source is configured to excite the long afterglow luminescent complex in the cuvette and is turned off after the excitation is completed, and the photon detector is configured to collect the luminescence signal emitted by the long afterglow luminescent complex after the excitation light source is turned off. The device directly detects the whole blood sample in the blood collection tube, and can realize fully automatic detection.

Description

Full-automatic immunoassay device

Technical Field

The disclosure belongs to the technical field of immunoassay, and particularly relates to a full-automatic immunoassay device.

Background

The detection of disease diagnosis is of great significance to human health and environmental safety, and is widely carried out in various medical sites. In the immune response for disease diagnosis and detection, the antigen can stimulate the immune system of animal body, induce immune response, and produce antibody with immune function in body fluid. The antibody and the antigen are combined through immunoreaction, the immunoreaction combination can be carried out in vivo and in vitro, and the antibody has the advantage of high specificity. Based on the derivation, immunoassay technology is derived and is applied to the field of biomedicine at present. Among the numerous immunoassay techniques, the luminescence immunoassay technique is performed by detecting a luminescence signal. With the great development of photon detection technology, the luminescence immunoassay technology can theoretically obtain higher detection sensitivity, and thus has received much attention.

The detection of biological samples such as blood is important in immunoassays. The traditional luminescence immunoassay detection has the problems of biological background interference and excitation light scattering, is difficult to directly detect the whole blood sample, and needs to pre-process the whole blood sample. However, the process of pre-processing of whole blood samples is cumbersome and presents a potential risk of human infection or sample contamination. For example, many of the conventional immunoassay devices perform detection using serum, and before detection, the cap of the blood collection tube is opened and the collected blood is centrifuged, so that it is not possible to directly detect a whole blood sample in the blood collection tube used daily in a hospital. The pretreatment process for serum detection can lead to a number of potential problems: high labor cost, error in manual operation, infection of medical staff by certain substances in blood, and the like.

In addition, most of the existing immunoassay instruments cannot continuously and automatically replace reaction cups in operation, and do not have the function of continuously detecting a large batch of whole blood samples, so that the high-throughput full-automatic detection of large batch continuity is difficult to realize.

The existing luminescence immunoassay instrument can not meet the detection requirement of full-automatic immunoassay of a whole blood sample, and also has the defects of complex structure, large volume, high cost, low test flux, less total continuous test amount, inconvenient operation and the like.

Disclosure of Invention

It is an object of the present disclosure to provide a fully automated immunoassay device that overcomes at least one of the deficiencies of the prior art.

The subject technology of the present disclosure is illustrated in accordance with aspects described below. For convenience, various examples of aspects of the subject technology are described as clauses (1, 2, 3, etc.) of the reference numerals. These terms are provided as examples and do not limit the subject technology of the present disclosure.

1. A fully automated immunoassay device, wherein the device is configured to perform an immunoassay test directly on a whole blood sample without pre-processing the whole blood sample, the device comprising:

the detection mechanism of the detection assembly comprises an excitation light source and a photon detector;

a sample supply assembly configured to provide a whole blood sample to be tested to the testing assembly in an automated manner;

a reagent supply assembly configured to provide one or more reagent components to a detection assembly in an automated manner;

a cuvette supply assembly configured to supply cuvettes to the detection assembly in an automated manner;

wherein the one or more reagent components and a target in the whole blood sample to be detected are immunoreactive in a reaction cup at the detection component to generate a long-lasting luminescent complex,

the excitation light source is configured to excite the long-afterglow luminescent compound in the reaction cup and is closed after excitation is completed, and the photon detector is configured to collect a luminescent signal emitted by the long-afterglow luminescent compound after the excitation light source is closed.

2. The fully automated immunoassay device of clause 1, wherein the one or more reagent components comprise: a first reagent component comprising a first antibody to the target and a second reagent component comprising a second antibody to the target, the first reagent component comprising one or more of a light absorbing agent, a buffering agent, and a light emitting agent, the second reagent component comprising the remaining of the light absorbing agent, the buffering agent, and the light emitting agent.

3. The fully automated immunoassay device of clause 2, wherein the one or more reagent components further comprises a third reagent component for diluting the whole blood sample.

4. The fully automated immunoassay device according to clause 1, wherein the one or more reagent components contain an additive component for hemolysis and/or signal amplification in an immunological reaction, and the additive component is selected from one or more of the group consisting of a hemolytic agent, salt, stabilizer, nanosphere, antibody, antigen, protein, surfactant, water, preservative, nucleic acid, polypeptide.

5. The fully automatic immunoassay device of any of clauses 1-4, wherein the detection mechanism further comprises a filter disposed between the excitation light source and the photon detector, the filter configured to filter out the excitation light when the excitation light source is turned on.

6. The fully automated immunoassay device according to any of clauses 1 to 4, wherein the detection mechanism is configured to obtain the concentration of the target in the whole blood sample from the intensity of the luminescence signal emitted from the photochemical long-afterglow luminescent complex according to a preset relationship curve between the intensity of the long-afterglow luminescence signal and the concentration of the target.

7. The fully automated immunoassay device of any of clauses 1-4, wherein the detection assembly further comprises an incubation mechanism adjacent to the detection mechanism, the incubation mechanism configured to incubate the one or more reagent components and the whole blood sample in the reaction cuvette to a specified temperature.

8. The fully automated immunoassay device of clause 7, wherein the incubation mechanism comprises an incubation tray provided with a plurality of reaction cup holders circumferentially spaced apart and receiving reaction cups.

9. The fully automated immunoassay device according to clause 7, wherein the detection assembly further comprises a moving gripper configured to move the cuvette between the incubation mechanism, the detection mechanism, and the cuvette feed assembly.

10. The fully automated immunoassay device according to any one of clauses 1 to 4, wherein the excitation light source comprises one or more selected from the group consisting of: solid laser, gas laser, semiconductor laser, photodiode, D65 standard light source, light emitting diode, ultraviolet lamp, xenon lamp, sodium lamp, mercury lamp, tungsten lamp, incandescent lamp, and fluorescent lamp.

11. The fully automatic immunoassay device according to any one of clauses 1 to 4, wherein the light wavelength of the excitation light source covers 300nm to 1000 nm.

12. The fully automatic immunoassay device according to any one of clauses 1 to 4, wherein the light wavelength of the excitation light source covers 600nm to 800 nm.

13. The fully automatic immunoassay device according to any one of clauses 1 to 4, wherein the excitation light source emits light in a focused beam, a divergent beam, an annular beam, or a collimated beam.

14. The fully automated immunoassay device of any of clauses 1-4, wherein the photon detector comprises one or more selected from the group consisting of: the device comprises a single photon counter, a photomultiplier, a silicon photocell, a photometric integrating sphere and a photographing imaging device.

15. The fully automated immunoassay device according to any one of clauses 1 to 4, wherein the reaction cup supply assembly comprises a cup magazine configured to receive the reaction cups to be used mixed out of order, and a sieve cup mechanism configured to automatically sequence the reaction cups in the cup magazine.

16. The fully automatic immunoassay device according to clause 15, wherein the reaction cuvette includes a cup body and a flange outwardly protruding from an upper portion of an outer surface of the cup body.

17. The fully automatic immunoassay device according to clause 16, wherein the sieve cup mechanism comprises an inclined guide tube having an inlet port at the upper outlet below, and an inclined parallel-bar chute having an inlet port at the upper outlet below, the inlet port of the inclined parallel-bar chute being located at the outlet port of the inclined guide tube, the inner diameter of the inclined guide tube being slightly larger than the outer diameter of the flange of the reaction cup, and the distance between the parallel bars of the inclined parallel-bar chute being larger than the outer diameter of the cup body of the reaction cup but smaller than the outer diameter of the flange.

18. The fully automated immunoassay device of clause 17, wherein the sieve cup mechanism further comprises a disposal tray at the exit of the parallel bar slide, the disposal tray configured to rotate to transport reaction cups off the inclined parallel bar slide.

19. The fully automated immunoassay device of clause 15, wherein the cup storage bin is funnel-shaped and its outlet leads to the sieve cup mechanism.

20. The fully automated immunoassay device according to any of clauses 1-4, wherein the reagent supply assembly comprises a reagent reservoir configured to contain the one or more reagent components, and a reagent transfer mechanism configured to transfer the one or more reagent components in the reagent reservoir into a reaction cup at the detection assembly.

21. The fully automated immunoassay device of clause 20, wherein the reagent reservoir comprises a plurality of compartments configured to contain the one or more reagent components.

22. The fully automated immunoassay device of clause 20, wherein the reagent transfer mechanism comprises a reagent needle configured to withdraw and release the one or more reagent components and a support arm supporting the reagent needle configured to move the reagent needle between the reagent reservoir and the reaction cup at the detection assembly.

23. The fully automated immunoassay device of clause 22, wherein the reagent needle is provided with a self-cleaning mechanism to prevent cross-contamination between the two samplings.

24. The fully automated immunoassay device according to any of clauses 1 to 4, wherein the specimen supply assembly comprises a sample introduction mechanism and a sampling mechanism, the sample introduction mechanism is configured to sequentially transport one or more blood collection tubes containing the whole blood specimen to the blood collection tube holder, and the sampling mechanism is configured to sample the blood collection tubes on the blood collection tube holder and deliver the whole blood specimen to the reaction cup at the test assembly.

25. The full-automatic immunoassay device according to clause 24, wherein the sample injection mechanism comprises a tube inlet bin, the blood collection tube seats and a tube outlet bin which are connected in series, the tube inlet bin is configured to receive a plurality of rows and a plurality of columns of blood collection tubes to be sampled, and the tube outlet bin is configured to receive a plurality of rows and a plurality of columns of sampled blood collection tubes.

26. The fully automated immunoassay device of clause 25, wherein the tube feeding magazine comprises a lateral pusher and/or a longitudinal pusher for pushing the blood collection tubes to the blood collection tube holders.

27. The fully automatic immunoassay device according to clause 25, wherein the cartridge is provided with a scanning mechanism for scanning the identification information on the cartridge.

28. The fully automated immunoassay device of clause 24, wherein the sampling mechanism comprises a guide rail located above the reaction cups of the sample introduction mechanism and the test assembly, a sample needle, and a drive mechanism that drives the sample needle to move on the guide rail between the lancet holder and the reaction cup at the test assembly.

29. The fully automated immunoassay device of clause 28, wherein the sample needle is provided with a self-cleaning mechanism to prevent cross-contamination between samples.

30. The fully automated immunoassay device of clause 24, wherein the specimen supply assembly further comprises a sample homogenizing mechanism for homogenizing the whole blood sample in the blood collection tube, the sample homogenizing mechanism being disposed adjacent to the blood collection tube holder.

31. The fully automated immunoassay device of clause 24, wherein the specimen supply assembly further comprises a washing mechanism configured to wash the sample needle and/or the reagent needle of the reagent supply assembly, the washing mechanism being located between the sample introduction mechanism and the detection assembly.

32. The fully automated immunoassay device of clause 31, wherein the wash mechanism comprises a wash cup and a wash line, and the wash line is configured to add a wash solution to the wash cup.

33. The fully automated immunoassay device of clause 32, wherein the wash cup is provided with a self-cleaning mechanism to prevent contamination of the sample needle and/or the reagent needle between washes.

34. The fully automatic immunoassay device according to clauses 1 to 4, wherein the detection module, the sample supply module, the reagent supply module and the cuvette supply module are provided on the same rack.

35. A method of testing a fully automated immunoassay device, wherein the device is configured to perform an immunoassay test directly on a whole blood sample without pre-processing the whole blood sample, the method comprising:

the cup screening mechanism receives a plurality of reaction cups which are mixed disorderly from the cup storage bin, and sequentially sorts the reaction cups in a cup mouth-up mode and sends the reaction cups to a cup outlet position;

moving the gripper to convey the sequenced reaction cups in the cup sieving mechanism to a reaction cup seat of the incubation mechanism from a cup outlet position;

the sample feeding mechanism receives one or more blood sampling tubes containing whole blood samples in the tube feeding bin and sequentially conveys the one or more blood sampling tubes to the blood sampling tube seats;

the sample is punctured and sampled aiming at a blood collection tube on a blood collection tube seat, and the collected whole blood sample is transferred into a reaction cup on a reaction cup seat;

the reagent needle adds one or more reagent components in different chambers of the reagent reservoir into the whole blood sample in the reaction cup on the reaction cup seat, wherein the one or more reagent components and the target object in the whole blood sample are subjected to immunoreaction in the reaction cup on the reaction cup seat to generate a long-afterglow luminescent complex;

the incubation mechanism incubates the one or more reagent components in the reaction cup and the whole blood sample at a set temperature and time; and

the movable gripper conveys the reaction cup from the incubation mechanism to the detection mechanism, an excitation light source of the detection mechanism excites the long-afterglow luminescent compound in the reaction cup and closes the reaction cup after excitation is finished, and the photon detector collects a long-afterglow luminescent signal emitted by the photochemical long-afterglow luminescent compound in the reaction cup.

36. The detecting method according to clause 35, wherein the detecting mechanism fits the intensity of the long-afterglow luminescence signal collected by the photon detector to a preset relation curve between the intensity of the long-afterglow luminescence signal and the concentration of the target object, so as to obtain the concentration of the target object in the whole blood sample.

37. The testing method of clause 35, wherein the gripper of the sample mixing mechanism grips the blood collection tube on the blood collection tube holder and mixes the whole blood sample in the blood collection tube, and then returns the whole blood sample to the blood collection tube holder.

38. The testing method of clause 35, wherein after the sample is punctured and sampled against the blood collection tube on the blood collection tube holder, the sample needle transfers the collected whole blood sample into the reaction cup, and the reagent needle extracts a reagent component of the one or more reagent components to dilute the whole blood sample in the reaction cup.

39. The assay of clause 35, wherein the device is a fully automated immunoassay device according to any one of clauses 1-34.

Additional features and advantages of the disclosed subject technology will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the disclosed subject technology. The advantages of the subject technology of the present disclosure will be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the subject technology of the present disclosure as claimed.

Drawings

Various aspects of the disclosure will be better understood upon reading the following detailed description in conjunction with the drawings in which:

fig. 1-4 show a top view, a front view, two side views of a fully automated immunoassay device according to embodiments of the present disclosure;

FIGS. 5 and 6 show front and top views of a sample injection mechanism of the fully automated immunoassay device of FIG. 1;

FIG. 7 illustrates a front view of the sampling mechanism of the fully automated immunoassay device of FIG. 1;

FIGS. 8 and 9 show front and side views of a sample mixing mechanism of the fully automated immunoassay device of FIG. 1;

FIGS. 10 and 11 show two side views of the washing mechanism and the rack of the fully automated immunoassay device of FIG. 1;

FIGS. 12 and 13 show top and front views of a reagent reservoir of the fully automated immunoassay device of FIG. 1;

FIGS. 14 and 15 show front and side views of a reagent transfer mechanism of the fully automated immunoassay device of FIG. 1;

FIGS. 16 and 17 show front and side views of a sieve cup mechanism of the fully automated immunoassay device of FIG. 1;

FIGS. 18 and 19 show top and front views of an incubation mechanism of the fully automated immunoassay device of FIG. 1;

FIG. 20 shows a perspective view of a moving hand grip of the fully automated immunoassay device of FIG. 1;

FIGS. 21 and 22 show front and side views of a detection mechanism of the fully automated immunoassay device of FIG. 1;

fig. 23 shows an operation flowchart of the fully automatic immunoassay device of fig. 1.

Detailed Description

The present disclosure will now be described with reference to the accompanying drawings, which illustrate several embodiments of the disclosure. It should be understood, however, that the present disclosure may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, the embodiments described below are intended to provide a more complete disclosure of the present disclosure, and to fully convey the scope of the disclosure to those skilled in the art. It is also to be understood that the embodiments disclosed herein can be combined in various ways to provide further additional embodiments.

It should be understood that like reference numerals refer to like elements throughout the several views. In the drawings, the size of some of the features may be varied for clarity.

It is to be understood that the terminology used in the description is for the purpose of describing particular embodiments only, and is not intended to be limiting of the disclosure. All terms (including technical and scientific terms) used in the specification have the meaning commonly understood by one of ordinary skill in the art unless otherwise defined. Well-known functions or constructions may not be described in detail for brevity and/or clarity.

As used in this specification, the singular forms "a", "an" and "the" include plural referents unless the content clearly dictates otherwise. The terms "comprising," "including," and "containing" when used in this specification specify the presence of stated features, but do not preclude the presence or addition of one or more other features. The term "and/or" as used in this specification includes any and all combinations of one or more of the associated listed items. The terms "between X and Y" and "between about X and Y" as used in the specification should be construed to include X and Y. The term "between about X and Y" as used herein means "between about X and about Y" and the term "from about X to Y" as used herein means "from about X to about Y".

In the description, when an element is referred to as being "on," "attached" to, "connected" to, "coupled" to, or "contacting" another element, etc., another element may be directly on, attached to, connected to, coupled to, or contacting the other element, or intervening elements may be present. In contrast, when an element is referred to as being "directly on," "directly attached to," directly connected to, "directly coupled to," or "directly contacting" another element, there are no intervening elements present. In the description, one feature is disposed "adjacent" another feature, and may mean that one feature has a portion overlapping with or above or below an adjacent feature.

In the specification, spatial relations such as "upper", "lower", "left", "right", "front", "rear", "high", "low", and the like may explain the relation of one feature to another feature in the drawings. It will be understood that the spatial relationship terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, features originally described as "below" other features may be described as "above" other features when the device in the figures is inverted. The device may also be otherwise oriented (rotated 90 degrees or at other orientations) and the relative spatial relationships may be interpreted accordingly.

The fully automatic immunoassay device according to the present disclosure performs immunoassay detection using the characteristics of the long persistence luminescent material. Long persistence phosphors are a special class of phosphors that can sustain luminescence for a period of time after the excitation light source is removed. The long-afterglow luminescent material has a luminescent life of more than one hundred milliseconds (even reaching the level of seconds or more) and has important application value in the fields of biomedicine, life science and the like. Unlike the existing inorganic long-afterglow luminescent materials based on photophysical processes, the newly developed long-afterglow luminescent materials suitable for immunoassay detection are based on organic systems. The organic system utilizes the characteristic of photochemical reaction, introduces photochemical reaction between light energy input and light energy output, and organically fuses photophysics and photochemistry together. In the long-afterglow luminescent material based on the organic system, a luminescent process relates to photochemical interaction among a plurality of chemical substances, wherein input excitation light energy is released in a luminescent form finally through a series of photochemical energy conversion and metabolic processes, so that long-afterglow luminescence is realized. Photochemical energy conversion and metabolism processes include energy input, energy buffering, energy extraction, energy transfer, and energy release. Originally, the very rapid photon radiation transition process (nanosecond magnitude to microsecond magnitude) is changed, energy is slowly released and is finally emitted in the form of light energy, so that the ultra-long light-emitting time (millisecond magnitude to hour magnitude) is obtained, the limitation of short light-emitting life of organic molecules is greatly improved, and the long-afterglow light-emitting intensity is improved.

The fully automatic immunoassay device according to the present disclosure improves the luminescent system of the photochemical long-afterglow luminescent material. The photochemical long afterglow luminescent material introduces photochemical reaction between light energy input and light energy output to slowly release photon radiation transition energy and finally emit the light energy, thereby obtaining ultra-long luminescence time, greatly improving the long afterglow luminescent performance and avoiding quenching caused by oxygen, water and the like. The photochemical long-afterglow luminescent material comprises a light absorbing agent, a buffering agent and a luminescent agent. The light absorbing agent, the buffering agent and the luminous agent can be randomly placed in various reagent components. The reagent and a target object (such as an antigen) in a blood sample to be detected form an immunological binding complex, the immunological binding complex forms a photochemical long-afterglow luminescent complex, and the immunological binding complex is used as a signal indicating probe to be applied to immunoassay detection, so that the interference of excitation light and background fluorescence is avoided, and the full-automatic washing-free homogeneous detection is realized.

Fig. 1 to 4 respectively show various angle views of a fully automated immunoassay device 1 according to an embodiment of the present disclosure. The fully automatic immunoassay device 1 directly detects the concentration of a target substance in a whole blood sample in a fully automatic manner. As shown in the drawings, the fully automatic immunoassay device 1 includes a sample supply module 10, a reagent supply module 20, a cuvette supply module 30, and a detection module 40, which are disposed on a holder 50. The sample supply assembly 10 is used to provide a whole blood sample to be tested to the testing assembly 40. The reagent supply assembly 20 is used to provide one or more reagent components to the detection assembly 40. The cuvette supply assembly 30 is used to provide cuvettes containing the whole blood sample and reagent components to the testing assembly 40. The test assembly 40 is used for performing an immunoassay test on a whole blood sample in a cuvette.

The sample supply assembly 10 includes a sample introduction mechanism 11 and a sampling mechanism 12. The sample injection mechanism 11 sequentially transfers one or more blood collection tubes 13 to the blood collection tube holder 112, and the sampling mechanism 12 samples the blood collection tubes 13 on the blood collection tube holder 112 and sends a whole blood sample to the detection module 40. As shown in fig. 5 and 6, the sample injection mechanism 11 includes a tube inlet bin 111, a blood collection tube seat 112, and a tube outlet bin 113 connected in series with each other. The tube feeding bin 111 can receive multiple rows and multiple columns of blood collection tubes 13 to be sampled, and each blood collection tube 13 to be sampled can hold a whole blood sample. The tube feeding bin 111 includes a transverse pushing member and a longitudinal pushing member, the transverse pushing member sequentially pushes the multiple blood collection tubes 13 to be sampled in each row to a sampling row aligned with the blood collection tube seat 112, and the longitudinal pushing member sequentially pushes the multiple blood collection tubes 13 to be sampled in the sampling row to the blood collection tube seat 112. The blood collection tube 13 on the blood collection tube seat 112 is sampled by the sampling mechanism 12 and then pushed to the tube outlet bin 113. The output tube bin 113 can receive a plurality of rows and columns of the sampled blood collection tubes 13. The outlet tube bin 113 includes a lateral pushing member and pushes the sampled blood collection tubes 13 located in the sampling row out of the sampling row. In some embodiments, the blood collection tube holder 112 may be provided with a scanning mechanism for scanning the identification information on the blood collection tube 13, thereby obtaining the traceability information of the whole blood sample inside the blood collection tube 13.

As shown in fig. 7, the sampling mechanism 12 includes a guide rail 121, a sample needle 122, and a driving mechanism 123 located above the sample introduction mechanism 11 and an incubation mechanism 41 (described in detail below) of the detection assembly 40. The driving mechanism 123 drives the sample needle 122 to move on the guide rail 121 to switch between a plurality of different positions (including a lancet holder position, a reaction cup holder position, and the like). For example, the driving mechanism 123 may drive the sample needle 122 to move to the lancet holder position, so that the sample needle 122 punctures and samples the blood collection tube 13 on the lancet holder 112. The driving mechanism 123 can also drive the sample needle 122 to move to the reaction cup seating position of the detecting assembly 40 to transfer the whole blood sample collected by the sample needle 122 into the reaction cup.

In some embodiments, the sample supply assembly 10 can further include a sample mixing mechanism 14 for mixing the sample in the blood collection tube 13, as shown in fig. 8 and 9. The sample mixing mechanism 14 is disposed adjacent to the lancet holder 112 and includes a grip 141. The grip 141 can grip the blood collection tube 13 on the blood collection tube holder 112 and mix the whole blood sample in the blood collection tube 13, and then return the whole blood sample to the blood collection tube holder 112.

In some embodiments, sample supply assembly 10 can further include a cleaning mechanism 15 for cleaning sample needle 122, as shown in fig. 10 and 11. The cleaning mechanism 15 is located between the sample injection mechanism 11 and the detection assembly 40. The cleaning mechanism 15 includes a cleaning cup 151 and a cleaning line 152, and the cleaning line 152 is used to add a cleaning liquid into the cleaning cup 151. The sample needle 122 is driven by the driving mechanism 123 to transfer the whole blood sample collected from the blood collection tube 13 into the reaction cup at the reaction cup holder position of the test assembly 40, and then the sample needle 122 is moved into the cleaning cup 151 for cleaning, thereby preventing cross contamination between two samplings. In some embodiments, the wash cup 151 may be provided with a self-cleaning mechanism (e.g., washing with a wash solution configured with a strong acid or strong base) to prevent contamination of the sample needle 122 between washes. In some embodiments, the sample needle 122 may be provided with a self-cleaning mechanism (e.g., cleaning with a cleaning solution configured with a strong acid or strong base).

The reagent supply assembly 20 includes a reagent reservoir 21 and a reagent transfer mechanism 22. The reagent reservoir 21 contains a reagent component that immunoreacts with a target in the whole blood sample, and the reagent transfer mechanism 22 transfers the reagent component in the reagent reservoir 21 into a reaction cup at the reaction cup holder position of the test assembly 40. As shown in fig. 12 and 13, the reagent reservoir 21 comprises a plurality of compartments for containing the reagent components, the temperature within the compartments being maintained at approximately 4 ℃ to approximately 8 ℃. The compartments respectively contain one or more reagent components, such as light absorbers, buffers, luminescent agents, etc. for achieving photochemical long afterglow luminescence. The reagent component is capable of immunoreacting with a target in a whole blood sample to be detected and coupling via immunoreaction to form an immunological binding complex, thereby producing a photochemical long-lasting luminescent signal detectable by the detection assembly 40.

In some embodiments, the light absorbing agent is disposed in the R1 reagent component, and the buffering agent and the light emitting agent are disposed in the R2 reagent component, wherein the R1 reagent component comprises a first antibody that immunoreacts with a target; the R2 reagent component contains a second antibody that immunoreacts with the target. That is, the R1 reagent component and the R2 reagent component contain components necessary for the photochemical long afterglow luminescence function, such as a light absorbent, a buffering agent, and a luminescent agent, and the R1 reagent component contains a first antibody to a target, and a photochemical long afterglow light absorbent; the R2 reagent component contains a secondary antibody of a target object, and a buffering agent and a luminous agent of photochemical long afterglow. The reagent components R1 and R2 also contain additives commonly used in immunological reactions for hemolysis and/or signal amplification, such as hemolytic agents, salts, stabilizers, signal amplification components, nanospheres, antibodies, antigens, proteins, surfactants, water, preservatives, nucleic acids, polypeptides, and the like. Some embodiments further include an R3 reagent component for diluting the whole blood sample, the R3 reagent component may be, for example, at least one of PB, PBs, PBST, BBS, MES, Tris, TES, HEPES, and the above additive components commonly used in immune reactions are also included in the R3 reagent component.

As shown in fig. 14 and 15, the reagent transfer mechanism 22 includes a reagent needle 221 and a support arm 222 that supports the reagent needle 221. The reagent needle 221 may extract and release a reagent component. The support arm 222 may move the reagent needle 221 in vertical and horizontal directions to switch between a plurality of different positions (e.g., reagent position, reaction cup position, etc.). For example, the support arm 222 may move the reagent needle 221 to reagent locations of multiple chambers of the reagent reservoir 21 to collect respective reagent components. The support arm 222 may move the reagent needle 221 to a reaction cup seating position of the detecting assembly 40 to transfer the reagent components collected by the reagent needle 221 into the reaction cup. In some embodiments, the support arm 222 is in the shape of an inverted L and includes a horizontal arm and a vertical arm. The free end of the horizontal arm is provided with a reagent needle 221 perpendicular thereto, and the vertical arm is rotated by a motor to move the reagent needle 221 between the reagent positions of the plurality of chambers of the reagent reservoir 21 and the reaction cup position of the detecting assembly 40.

The reagent needle 221 may also be provided with a reagent needle self-cleaning mechanism 223 (e.g., cleaning with a cleaning solution configured with an acid, base, and/or surfactant) as shown in fig. 1 to prevent cross-contamination between two samplings. In some embodiments, the reagent needle 221 may be moved into the washing mechanism 15 for washing.

The reaction cup supply assembly 30 includes a cup magazine and a sieve cup mechanism 32. The cup storage bin is used for receiving reaction cups to be used, and the cup screening mechanism 32 is used for sequencing the reaction cups which are mixed in disorder in the cup storage bin. The reaction cup comprises a cup body and a flange extending outwards from the upper part of the outer surface of the cup body. The cup magazine is funnel shaped and its outlet is located at the inlet of the sieve cup mechanism 32. As shown in fig. 16 and 17, the sieve cup mechanism 32 includes a guide tube 321, a parallel bar chute 322, and a disposal tray 323. The guide pipe 321 is placed obliquely with the inlet at the upper outlet and the inner diameter slightly larger than the outer diameter of the flange of the reaction cup. The reaction cup can be transported in the guide pipe 321 with its mouth facing either upward or downward. The parallel-bar chute 322 is disposed obliquely with its inlet at the upper outlet and its inlet at the outlet of the guide pipe 321. The distance between the parallel bars of the parallel bar slideway 322 is larger than the outer diameter of the cup body of the reaction cup but smaller than the outer diameter of the flange. The reaction cup leaves the guide pipe 321, slides on the parallel-bar slideway 322 by the extended flange thereof, and the cup mouth is automatically turned upwards under the action of gravity. A disposal tray 323 is located at the exit of the parallel bar chute 322 and can rotate to transport reaction cups exiting the parallel bar chute 322 to a cup exit position.

The detection assembly 40 comprises an incubation mechanism 41 and a detection mechanism 42. The incubation mechanism 41 is used to incubate the whole blood sample in the cuvette to a specified temperature (e.g., 37.5 ℃), and the detection mechanism 42 is used to detect the optical signal generated by the whole blood sample in the cuvette. As shown in fig. 18 and 19, the incubation mechanism 41 includes an incubation tray 411, and the incubation tray 411 is provided with a plurality of reaction cup holders 412 spaced apart in the circumferential direction to receive reaction cups. The plurality of reaction cup holders 412 are rotatable about a central axis of the incubation mechanism 41. Moving the gripper 43 (as shown in fig. 20) sequentially transfers the cuvettes at the cup exit position of the cuvette supply assembly 30 into the cuvette holder 412, and the sample needle 122 and the reagent needle 221 transfer the diluted whole blood sample and the reagent components into the cuvettes, respectively. The movable gripper 43 can pick up and mix the reaction cups from the reaction cup holder 412, and then return the reaction cups to the reaction cup holder 412 or convey the reaction cups to the detection mechanism 42.

As shown in FIGS. 21 and 22, the detection mechanism 42 is disposed adjacent to the incubation mechanism 41. The detection mechanism 42 includes an excitation light source, a photon detector, and a filter disposed therebetween. The excitation light source is used to excite the photochemical long persistence luminescent complex in the whole blood sample and is turned off after excitation is complete. The filter is used for filtering exciting light and protecting the photon detector. The photon detector is used for collecting the luminescent signal emitted by the photochemical long afterglow luminescent compound.

As described above, the R1 reagent component, R2 reagent component and the target in the whole blood sample form an immunological binding complex in the cuvette. The immunological combination compound forms a photochemical long-afterglow luminescent compound, and the intensity of the long-afterglow luminescent signal of the photochemical long-afterglow luminescent compound has positive correlation with the concentration of a target substance in a whole blood sample. The detection mechanism stores a preset relation curve between the intensity of the long afterglow luminescence signal and the concentration of the target object. The detection mechanism 42 can detect the concentration of the target object according to the intensity of the photochemical long afterglow luminescence signal collected by the photon detector.

In some embodiments, the excitation light source may be a solid-state laser, a gas laser, a semiconductor laser, a photodiode, a D65 standard light source, a light emitting diode, an ultraviolet lamp, a xenon lamp, a sodium lamp, a mercury lamp, a tungsten lamp, an incandescent lamp, a fluorescent lamp, and combinations thereof. In some embodiments, the excitation light source may be a laser or a light emitting diode, and these light sources have good monochromaticity and high luminance of output light, and can selectively and rapidly excite energy charging. The light emitted by the excitation light source may be a focused, divergent, annular, collimated beam. The wavelength of the excitation light source may cover 300nm-1000nm, more particularly 600 nm-800 nm. For example, a photodiode can be used as an excitation light source, the light wavelength of the photodiode is about 730nm, the photodiode is switched off after excitation for 1 second, and the photochemical long afterglow luminescent compound in the reaction cup then emits an upconversion type long afterglow luminescent signal about 610 nm.

In some embodiments, the photon detector may be a single photon counter, a photomultiplier tube, a silicon photocell, a photometric integrating sphere, or a photographic imaging device.

The sample detection step of the fully automatic immunoassay device 1 according to the embodiment of the present disclosure is described below with reference to fig. 23. The sifter cup mechanism 32 of the reaction cup supply assembly 30 receives the unordered mixed reaction cups from the cup magazine. The reaction cups sequentially pass through the guide pipe 321, the parallel-bar chute 322 and the sorting tray 323 of the cup passing mechanism 32, and are sequentially sorted with the cup mouths facing upward and reach the cup discharge position.

The moving gripper 43 of the detecting assembly 40 transports the sequenced reaction cups in the sifter cup mechanism 32 from the cup exit position to the reaction cup holder 412 of the incubation mechanism 41.

The tube feeding bin 111 of the sample feeding mechanism 11 of the specimen supply assembly 10 sequentially transfers the received one or more blood collection tubes 13 containing the whole blood specimen to the blood collection tube seat 112 by using the lateral pushing member and the longitudinal pushing member thereof.

The grip 141 of the sample mixing mechanism 14 picks up the blood collection tube 13 on the blood collection tube holder 112, mixes the whole blood sample in the blood collection tube 13, and then returns the mixture to the blood collection tube holder 112.

The driving mechanism 123 of the sampling mechanism 12 of the specimen supply assembly 10 drives the specimen needle 122 to move on the guide rail 121 to the lancet holder position. The sample needle 122 punctures and samples the blood collection tube 13 on the blood collection tube holder 112, and is driven by the driving mechanism 123 to transfer a collected whole blood sample (for example, 10 μ l blood sample) into the reaction cup at the reaction cup holder 412. Then, the sample needle 122 is moved into the washing cup 151 for washing.

The reagent needle 221 of the reagent supply assembly 20 adds the R3 reagent component in the reagent reservoir 21 to the whole blood sample in the reaction cup to dilute the whole blood sample. After the dilution is performed, the reagent needle 221 is moved to the washing cup 151 for washing.

The reagent needle 221 adds the R1 reagent component and the R2 reagent component in the reagent reservoir 21 to the diluted whole blood sample in the reaction cup.

The moving grip 43 picks up the reaction cup and mixes the reagent and diluted whole blood sample in situ with high speed rotation, and then places it back into the reaction cup holder 412 of the incubation mechanism 41.

The incubation plate 411 of the incubation mechanism 41 is rotated to perform a set temperature and time incubation of the reagents in the reaction cup and the diluted whole blood sample.

After the incubation is completed, the hand grip 43 is moved to transfer the cuvette to the detection mechanism 42. An excitation light source (e.g., a photodiode with a luminescence wavelength of about 730 nm) of the detection mechanism 42 excites the reaction cup for several seconds (e.g., 1 second) and is turned off, and the photochemical long-afterglow luminescent compound in the reaction cup emits a long-afterglow luminescent signal (e.g., a red long-afterglow luminescent signal of about 610 nm). The detection mechanism 42 performs fitting according to the intensity of the long afterglow luminescence signal collected by the photon detector and a preset relation curve between the stored intensity of the long afterglow luminescence signal and the concentration of the target object, so as to obtain the concentration of the target object in the whole blood sample. After the detection is completed, the reaction cup is discarded.

According to this full-automatic immunoassay device of this disclosed embodiment can directly test whole blood sample, need not heparin tube and uncap the operation, also need not blood centrifugation operation. The target (e.g., antigen) is present in a whole blood sample, and the whole blood sample is directly collected, stored, and loaded by a blood collection tube commonly used in medical sites. Therefore, the fully automatic immunoassay device can directly perform a series of detection operations on a whole blood sample in a blood collection tube, and has a function of continuous batch detection.

The full-automatic immunoassay device according to the embodiment of the disclosure adopts a long afterglow luminescence detection technology to detect a whole blood sample. Because only the immunoconjugates can emit light signals after the exciting light is turned off, the intensity of the light signals corresponds to the concentration of the substance to be detected. The long afterglow luminescence detection technology can avoid the signal interference of background light and a light source and obtain a high-quality detection result.

According to the full-automatic immunoassay device disclosed by the embodiment of the disclosure, full-automatic immunoassay detection can be realized, and the labor amount of medical staff is effectively saved.

A fully automatic immunoassay device according to embodiments of the present disclosure uses a sieve cup mechanism. As long as reaction cups are poured into the cup storage bin, the cup sieving mechanism can upwards sort the orderly cup mouths of the reaction cups which are mixed disorderly. Compared with the prior art in which the well-sequenced reaction cups are manually placed, the cup sieving mechanism realizes the full-automatic sequencing of the reaction cups. As long as the reaction cups in the cup storage bin are replenished regularly, the full-automatic immunoassay device according to the embodiment of the disclosure can realize uninterrupted detection.

According to the full-automatic immunoassay device disclosed by the embodiment of the disclosure, the disposable reaction cup is used, the possibility of cross contamination of front and back tests is reduced, and a plurality of analysis tests can be simultaneously carried out. This design greatly increases the accuracy and repeatability of the test (since a disposable cuvette is used, the background of the photochemistry signal does not rise after multiple tests), and improves the efficiency of the instrument test (e.g., up to 180 tests per hour). The population of China is large, medical resources are scarce, and the full-automatic immunoassay device according to the embodiment of the disclosure can adapt to the requirements of longer detection time, more projects and heavier tasks of the existing detection projects to a greater extent.

According to the full-automatic immunoassay device disclosed by the embodiment of the disclosure, the full-automatic immunoassay device is formed by simplified modular components, the size is in a desktop level, and the defects of large occupied space and inconvenience in movement are avoided. Compared with the existing device, the full-automatic immunoassay device has the advantages of low test cost, quick linkage operation and convenience in test.

Although exemplary embodiments of the present disclosure have been described, it will be understood by those skilled in the art that various changes and modifications can be made to the exemplary embodiments of the present disclosure without substantially departing from the spirit and scope of the present disclosure. Accordingly, all changes and modifications are intended to be included within the scope of the present disclosure as defined in the appended claims. The disclosure is defined by the following claims, with equivalents of the claims to be included therein.

Claims

1. A fully automatic immunoassay device, characterized in that the device is configured to directly perform immunoassay detection on the whole blood sample without preprocessing the whole blood sample, the device comprising:

a detection assembly, the detection mechanism of the detection assembly includes an excitation light source and a photon detector;

a sample supply assembly configured to provide a whole blood sample to be tested to the detection assembly in an automated manner;

a reagent supply assembly configured to provide one or more reagent components to the detection assembly in an automated manner;

a cuvette supply assembly configured to provide cuvettes to the detection assembly in an automated manner;

wherein, the one or more reagent components and the target in the whole blood sample to be detected undergo an immune reaction in the cuvette at the detection component to generate a long afterglow luminescence complex,

Wherein, the excitation light source is configured to excite the long afterglow luminescent complex in the cuvette and is turned off after the excitation is completed, and the photon detector is configured to collect the luminescent signal emitted by the long afterglow luminescent complex after the excitation light source is turned off.

2 . The fully automatic immunoassay device according to claim 1 , wherein the one or more reagent components comprise: a first reagent component of a first antibody containing a target substance and a first reagent component of a first antibody containing the target substance. 3 . The second reagent component of the diabody, the first reagent component includes one or more of a light absorbing agent, a buffering agent, and a luminescent agent, and the second reagent component includes the remainder of the light absorbing agent, the buffering agent, and the luminescent agent kind.

3. The fully automatic immunoassay device of claim 2, wherein the one or more reagent components further comprise a third reagent component for diluting the whole blood sample.

4 . The fully automatic immunoassay device according to claim 1 , wherein the one or more reagent components contain additive components for hemolysis and/or signal amplification in the immune reaction, and the additive components One or more selected from the group consisting of hemolytic agents, salts, stabilizers, nanospheres, antibodies, antigens, proteins, surfactants, water, preservatives, nucleic acids, polypeptides.

5. The fully automatic immunoassay device according to any one of claims 1 to 4, wherein the detection mechanism further comprises a filter plate arranged between the excitation light source and the photon detector, and the filter plate is configured to Excitation light is filtered out when the excitation light source is turned on.

6. The fully automatic immunoassay device according to any one of claims 1 to 4, wherein the detection mechanism is configured to change from the photochemical long-term luminescence signal intensity to the target substance concentration according to a preset relationship curve The intensity of the luminescent signal emitted by the afterglow luminescent complex yields the concentration of the target in the whole blood sample.

7. The fully automatic immunoassay device according to any one of claims 1-4, wherein the detection assembly further comprises an incubation mechanism adjacent to the detection mechanism, wherein the incubation mechanism is configured to The one or more reagent components and the whole blood sample are incubated to the specified temperature.

8 . The fully automatic immunoassay device of claim 7 , wherein the incubation mechanism comprises an incubation plate provided with a plurality of cuvette holders spaced circumferentially and receiving cuvette. 9 .

9. The fully automatic immunoassay device of claim 7, wherein the detection assembly further comprises a moving gripper configured to move the cuvette between the incubation mechanism, the detection mechanism and the cuvette supply assembly.

10. The fully automatic immunoassay device according to any one of claims 1-4, wherein the excitation light source comprises one or more selected from the group consisting of: solid-state laser, gas laser, semiconductor laser, and photodiode , D65 standard light source, light-emitting diode, ultraviolet lamp, xenon lamp, sodium lamp, mercury lamp, tungsten lamp, incandescent lamp, fluorescent lamp.