Immunochromatographic analysis method, storage medium, and analyzer

Abstract

The present application discloses an immunochromatographic analysis method, a storage medium, and an analyzer. The analyzer comprises a sample collection device, a reagent supply device, a sample transfer device, a light source assembly, an optical measurement device comprising an optical imaging unit and an optical scanning unit, and a data processing unit. The data processing unit uses at least part of information in an acquired two-dimensional fluorescence signal image and / or at least part of information in a one-dimensional fluorescence signal curve to correct a measurement result to obtain a corrected measurement result, thereby improving the accuracy of measurement results.

Description

An immunochromatographic analysis method, storage medium and analyzer

[0001] This application claims priority to Chinese patent applications No. 202411837083X and 2024118371993, filed in China on December 13, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0002] This application relates to the field of medical testing, specifically to an immunochromatographic analysis method, storage medium, and analyzer. Background Technology

[0003] Immunochromatography belongs to the field of point-of-care testing (POCT) and has advantages such as convenient and rapid detection, low overall cost, and the ability to be stored and transported at room temperature. In optically-based quantitative immunochromatography, light emitted from a light source illuminates the immunoassay strip. A detector then collects the fluorescence signals generated by the detection line (T line) and / or control line (C line) of the immunoassay strip after illumination. The collected fluorescence signals are then converted into electrical signals, the magnitude of which is related to the concentration of the analyte, thereby quantitatively detecting the concentration of the analyte.

[0004] Differences in the raw materials used in chromatographic test strips can lead to variations in physical characteristics such as pore size and porosity between different chromatographic reagent cards, with particularly significant batch-to-batch variations. This affects the chromatography speed and the final test results, resulting in inaccurate readings. Furthermore, variations in the manufacturing process of chromatographic test strips can cause differences in the amount of reagent embedded in the test lines, fixation strength, and uniformity of distribution, affecting the test results and leading to inaccurate results. Moreover, simple signal intensity analysis cannot reflect the absence / incompleteness of the light signal in the T line and / or control line (C line), nor can it reflect the uniformity of the light signal, let alone the speed of the chromatographic reaction. Traditional immunochromatographic analysis methods that only obtain the light signal intensity after the chromatographic reaction suffer from poor accuracy and repeatability in POCT test results.

[0005] Therefore, this application proposes a new immunochromatographic analyzer and analytical method that can effectively solve the problems existing in traditional immunochromatographic analysis. Summary of the Invention

[0006] To address the problems associated with traditional immunochromatographic analysis, this application provides an immunochromatographic analysis method, storage medium, and analyzer.

[0007] Firstly, an immunochromatographic analyzer is provided, comprising:

[0008] A sample collection device, at least for collecting samples used to prepare test specimens;

[0009] A reagent supply device provides reagents that can react with the sample and participate in the sample preparation process;

[0010] A sample transfer device configured to transfer the sample to a test strip of an immunochromatographic analysis reagent card;

[0011] A light source assembly is configured to illuminate the test strip. The light source assembly is used to excite a first fluorescent substance to produce a first fluorescence and to excite a second fluorescent substance to produce a second fluorescence. The first fluorescent substance includes a first internal reference, which is present at least on the detection line of the test strip.

[0012] An optical measurement device, comprising an optical imaging unit and an optical scanning unit, wherein the optical imaging unit is at least used to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip, and the optical scanning unit is at least used to acquire a one-dimensional fluorescence signal curve of the immunochromatographic test strip;

[0013] The data processing unit is configured to correct the detection result by using at least some information from the two-dimensional fluorescence signal image and / or at least some information from the one-dimensional fluorescence signal curve, so as to obtain the corrected detection result.

[0014] An immunochromatographic assay strip includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, which has a detection line. The detection line is coated with a trap and a first fluorescent substance. Immunochromatographic analysis directly and / or indirectly captures tracers through the trap on the detection line. The tracers generate a light signal, which is used to indicate at least the presence and / or content of the analyte. However, the amount of trap on the detection line affects the amount of tracers directly and / or indirectly captured, thus affecting the magnitude of the light signal generated by the tracers, and consequently the detection results. Therefore, it is necessary to accurately determine the amount of trap on the detection line, as well as the flow cytometry conditions. Furthermore, the uniformity of the trap on the detection line also affects the uniformity of the tracers, which in turn affects the uniformity of the light signal generated by the tracers, and consequently the detection results. Therefore, it is also necessary to accurately determine the uniformity of the trap on the detection line.

[0015] To this end, this application provides an optical measurement device including an optical imaging unit and an optical scanning unit. The optical imaging unit is used at least to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip, and / or the optical scanning unit is used at least to acquire a one-dimensional fluorescence signal curve of the immunochromatographic test strip. Then, the detection result is corrected using at least some information from the two-dimensional fluorescence signal image and / or at least some information from the one-dimensional fluorescence signal curve to obtain a corrected detection result. Specifically, at least one detection result correction parameter can be obtained first using at least some information from the two-dimensional fluorescence signal image and / or at least some information from the one-dimensional fluorescence signal curve, and the detection result is corrected using the at least one detection result correction parameter to obtain a corrected detection result, thereby improving the accuracy of the detection result. As an example, at least some information in the two-dimensional fluorescence signal image may be fluorescence intensity value information, fluorescence intensity uniformity information, fluorescence signal location information, fluorescence signal time information, etc.; at least some information in the one-dimensional fluorescence signal image may be fluorescence intensity value information, fluorescence signal location information, fluorescence signal time information, etc.

[0016] Typically, the trap and the first fluorescent substance are coated onto the detection line using the same or similar processes. Therefore, by detecting the fluorescence signal of the first fluorescent substance, the coating status of the trap on the detection line can be obtained, including the coating amount and coating uniformity. The fluorescent signal substance can be at least one of fluorescent microspheres, quantum dots, fluorescent proteins, or luciferin molecules. Further, the first fluorescent substance includes a first internal control, which is present at least on the detection line of the test strip.

[0017] In some alternative approaches, the trap and the first internal reference are independently coated on the detection line; or, the first internal reference is directly and / or indirectly bound to the trap. Therefore, the coating status of the trap on the detection line can be indirectly obtained by detecting the fluorescence signal of the first fluorescent substance (first internal reference) on the detection line.

[0018] When the first internal control is directly and / or indirectly combined with the target, the coating amount of the first internal control shows better consistency with the coating amount of the target; the coating uniformity of the first internal control also shows better consistency with the coating uniformity of the target. That is, using the first internal control to indicate the coating amount and / or coating uniformity of the target in the detection line is reliable; correspondingly, by acquiring a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence generated by the first internal control, and using at least some information in the two-dimensional fluorescence signal image and / or at least some information in the one-dimensional fluorescence signal curve to correct the detection results, the accuracy of the detection results can be improved.

[0019] Optionally, the trap is selected from at least one of avidin, streptavidin, anti-biotin antibody, anti-FITC antibody, or anti-DNP antibody; or, the trap is selected from at least one of antibody, antigen-binding fragment, aptamer, modified aptamer, aptamer, avidin, antigen, protein, polypeptide, multi-protein complex, exosome, microbial particle / fragment / disc, oligonucleotide, or low molecular weight compound.

[0020] Furthermore, the immunochromatographic analyzer also includes a control unit, which is at least used to control the optical measurement device to acquire a first one-dimensional fluorescence signal curve C1 of the first fluorescence before the detection line in the chromatographic reaction, and / or at least used to control the optical measurement device to acquire a first two-dimensional fluorescence signal image I1 of the first fluorescence before the detection line in the chromatographic reaction.

[0021] Optionally, the immunochromatographic analyzer further includes a control unit, which is at least used to control the optical measurement device to acquire the second one-dimensional fluorescence signal curve C2 of the first fluorescence after the chromatography reaction is completed, and / or at least used to control the optical measurement device to acquire the second two-dimensional fluorescence signal image I2 of the first fluorescence after the chromatography reaction is completed.

[0022] In this application, the first fluorescent substance includes a first internal reference. A control unit controls an optical measurement device to acquire a first one-dimensional fluorescence signal curve C1 of the first fluorescence up to the detection line. From the first one-dimensional fluorescence signal curve C1, information such as the peak intensity and / or peak area of ​​the fluorescence signal corresponding to the detection line can be obtained, thus determining the actual coating amount of the trap on the test strip's detection line. The control unit also controls an optical measurement device to acquire a first two-dimensional fluorescence signal curve I1 of the first fluorescence up to the detection line. From the first two-dimensional fluorescence signal curve I1, information such as the intensity and intensity distribution of the fluorescence signal corresponding to the detection line can be obtained, thus determining the uniformity of the actual coating amount of the trap on the test strip's detection line. Differences in the actual coating amount and / or the actual coating uniformity of the trap on the detection line will affect the detection results. Therefore, it is necessary to acquire the first one-dimensional fluorescence signal curve C1 and / or the first two-dimensional fluorescence signal curve I1 of the first fluorescence up to the detection line, and correct the detection results based on at least some of the information therein, to obtain a corrected detection result. Specifically, at least one detection result correction parameter can be obtained first based on at least some of the information, and then the detection result can be corrected using the at least one detection result correction parameter.

[0023] Similarly, by controlling the optical measurement device through the control unit to acquire the second one-dimensional fluorescence signal curve C2 and the second two-dimensional fluorescence signal curve I2 of the first fluorescence after the chromatographic reaction, the effective coating amount and uniformity of the captured material on the detection line of the test strip after the chromatographic reaction can be obtained. Since the captured material may be lost during the chromatographic reaction, the actual coating amount and actual coating uniformity of the captured material on the detection line before the detection line are reached are not equal to the effective coating amount and effective coating uniformity of the captured material on the detection line. Therefore, it is necessary to acquire the second one-dimensional fluorescence signal curve C2 and / or the second two-dimensional fluorescence signal curve I2 of the first fluorescence after the chromatographic reaction, and correct the detection results based on at least some of the information therein to obtain the corrected detection results. Specifically, at least one detection result correction parameter can be obtained first based on at least some of the information, and then the detection results can be corrected using the at least one detection result correction parameter.

[0024] Furthermore, due to the loss of the captured material, the tracer material it captured will also be carried away, resulting in tracer loss. This causes the amount of tracer material captured in the detection line area to be lower than the actual amount, leading to inaccurate detection results. Therefore, by combining at least one of C1 and I1 and at least one of C2 and I2, information on the loss of the captured material during the chromatography process on the detection line can be obtained (e.g., loss amount, loss rate, loss uniformity, etc.), which can better correct the detection results.

[0025] In some embodiments, the second fluorescent substance includes a second internal reference. Before the chromatographic reaction, the second internal reference is present in the sample and / or in the non-detection line region of the test strip. During the chromatographic reaction, the second internal reference precipitates towards the detection line along the chromatographic direction of the test strip. By controlling the optical measurement device to acquire a third two-dimensional fluorescence signal image I3 of the second fluorescence during the chromatographic reaction, the current position of the chromatographic reaction can be obtained, and thus the rate of the chromatographic reaction can be determined. At different chromatographic reaction rates, the amount of tracer captured by the trap will differ. Therefore, in this application, by acquiring a third two-dimensional fluorescence signal image I3 of the second fluorescence during the chromatographic reaction, which contains information related to the chromatographic reaction rate, and using at least some of the information in the third two-dimensional fluorescence signal image I3 to correct the detection results, the accuracy of the detection results can be improved.

[0026] Preferably, the optical measuring device is controlled to acquire a fourth two-dimensional fluorescence signal image I4 and a fifth two-dimensional fluorescence signal image I5 of the second fluorescence at two different moments during the chromatography reaction process. Based on the position of the current chromatography reaction in the two-dimensional fluorescence signal images at two different moments, information related to the chromatography reaction rate can be better reflected.

[0027] Furthermore, the optical measuring device also includes a first filter and a second filter, wherein the first filter is used to filter out the first fluorescence and the second filter is used to filter out the second fluorescence.

[0028] Furthermore, the optical imaging unit includes at least one of CCD and CMOS; the optical scanning unit includes a photodetector.

[0029] Furthermore, the immunochromatographic analyzer also includes an information acquisition unit, which is used to acquire reference information of the immunochromatographic analysis reagent card. The reference information includes at least the reference concentration information R0 of the first fluorescent substance on the detection line and / or the reference fluorescence signal T0 of the first fluorescent substance on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve.

[0030] Accordingly, the data processing unit is at least configured to combine at least some information in the reference information, at least some information in the two-dimensional fluorescence signal image and / or at least some information in the one-dimensional fluorescence signal curve to correct the detection result, thereby obtaining a corrected detection result.

[0031] Furthermore, the data processing unit is at least configured to combine at least some information in the reference information, at least some information in the two-dimensional fluorescence signal image, and / or at least some information in the one-dimensional fluorescence signal curve to obtain at least one detection result correction parameter, wherein the at least one detection result correction parameter is used to correct the detection result to obtain a corrected detection result.

[0032] The detection result correction parameters acquired by the data processing unit include at least the following:

[0033] The following are possible combinations of: a first capture coating amount correction factor related to the amount of capture on the detection line before the chromatographic reaction proceeds to the detection line; a second capture coating amount correction factor related to the amount of capture on the detection line after the chromatographic reaction is completed; a first capture coating uniformity correction factor related to the amount of capture on the detection line before the chromatographic reaction proceeds to the detection line; a second capture coating uniformity correction factor related to the amount of capture on the detection line after the chromatographic reaction is completed; and an actual chromatographic rate correction factor related to the actual chromatographic reaction rate.

[0034] Furthermore, the control unit is at least used to control the optical measuring device to acquire the third one-dimensional fluorescence signal curve C3 and / or the sixth two-dimensional fluorescence signal image I6 of the second fluorescence in the detection line region after the chromatography reaction is completed, and the third one-dimensional fluorescence signal curve C3 and / or the sixth two-dimensional fluorescence signal image I6 are at least used to acquire the detection result.

[0035] Furthermore, the immunochromatographic analyzer also includes a result output unit, which is used to output at least the detection result and / or at least one corrected detection result; preferably, it simultaneously outputs the detection result, at least one corrected detection result, and the correction parameter corresponding to the corrected detection result. Optionally, the result output unit is used to output at least the detection result and at least two corrected detection results for selection.

[0036] Preferably, the result output unit includes a display interface, which is used to display at least the detection result and / or at least one corrected detection result. More preferably, the display interface is used to display at least the detection result and at least two corrected detection results for selection, and in response to a selection command, outputs the selected detection result or the corrected detection result as the final detection result.

[0037] Furthermore, the light source assembly is at least configured to provide a first excitation light of a first wavelength and a second excitation light of a second wavelength, wherein the first excitation light is used to excite a first fluorescent substance to produce a first fluorescence, and the second excitation light is used to excite a second fluorescent substance to produce a second fluorescence. Alternatively, the light source assembly is at least configured to provide a third excitation light of a third wavelength, wherein the third excitation light is used to excite the first fluorescent substance to produce a first fluorescence, and the third excitation light is used to excite the second fluorescent substance to produce a second fluorescence.

[0038] Preferably, the data processing unit is further configured to convert the two-dimensional fluorescence signal image into a three-dimensional fluorescence signal image; the result output unit is further configured to output the one-dimensional fluorescence signal curve and / or the two-dimensional fluorescence signal image and / or the three-dimensional fluorescence signal image.

[0039] Furthermore, the immunochromatographic analyzer also includes a sample loading module for automatically loading and unloading sample holders or sample tubes; and / or a reagent card loading module for automatically loading reagent cards; and / or an incubation module for carrying and incubating reagent cards; and / or a temperature control unit for providing the temperature conditions required for incubation; and / or a reagent card unloading module for automatically unloading reagent cards.

[0040] Optionally, the test strip further includes a conjugate pad located between the sample pad and the NC membrane. Alternatively, the test strip may not have a conjugate pad, and the sample pad may be directly connected to the NC membrane.

[0041] Secondly, an immunochromatographic analysis method is provided, including:

[0042] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. The immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least a first internal control and a trap. The trap and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the trap.

[0043] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0044] The sample is fed to the test strip of the immunochromatographic assay reagent card to initiate the chromatographic reaction; the sample contains a tracer;

[0045] The reagent card is illuminated by a light source, which excites the tracer to produce a second fluorescence;

[0046] Acquire the signal of the first fluorescence; the signal of the first fluorescence includes: a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the immunochromatographic analysis reagent card is provided to the immunochromatographic analyzer and the chromatography reaction proceeds to the detection line; and a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the chromatography reaction is completed;

[0047] Acquire the signal of the second fluorescence; the signal of the second fluorescence includes: a two-dimensional fluorescence signal image of the second fluorescence and / or a one-dimensional fluorescence signal curve of the second fluorescence after the chromatography reaction is completed;

[0048] The detection result is output by combining at least a portion of the information from the first fluorescence signal and at least a portion of the information from the second fluorescence signal.

[0049] Furthermore, the immunochromatographic analysis method further includes:

[0050] Obtain reference information for the immunochromatographic assay reagent card, the reference information including at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve;

[0051] The detection result is output by combining at least a portion of the information from the first fluorescence signal, at least a portion of the information from the second fluorescence signal, and at least a portion of the information from the reference information.

[0052] Thirdly, an immunochromatographic analysis method is provided, including:

[0053] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. The immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least a first internal control and a trap. The trap and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the trap.

[0054] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0055] The sample is fed to the test strip of the immunochromatographic assay reagent card to initiate the chromatographic reaction; the sample contains a tracer;

[0056] The reagent card is illuminated by a light source, which excites the tracer to produce a second fluorescence;

[0057] Acquire the signal of the first fluorescence; the signal of the first fluorescence includes: a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the immunochromatographic analysis reagent card is provided to the immunochromatographic analyzer and the chromatography reaction proceeds to the detection line and / or after the chromatography reaction is completed;

[0058] Acquire the signal of the second fluorescence; the signal of the second fluorescence includes: a two-dimensional fluorescence signal image of the second fluorescence and / or a one-dimensional fluorescence signal curve of the second fluorescence after the chromatography reaction is completed, and a two-dimensional fluorescence signal image of the second fluorescence during the chromatography reaction.

[0059] The detection result is output by combining at least a portion of the information from the first fluorescence signal and at least a portion of the information from the second fluorescence signal.

[0060] Furthermore, the immunochromatographic analysis method further includes:

[0061] Obtain reference information for the immunochromatographic assay reagent card, the reference information including at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve;

[0062] The detection result is output by combining at least a portion of the information from the first fluorescence signal, at least a portion of the information from the second fluorescence signal, and at least a portion of the information from the reference information.

[0063] Fourthly, an immunochromatographic analysis method is provided, comprising:

[0064] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. The immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and the NC membrane has a detection line. The detection line is at least coated with a trapping agent. The trapping agent is used to capture tracers during the chromatography process.

[0065] Provide a sample to a reagent card, the sample containing a tracer, such that the tracer in the sample is chromatographically deposited on the test strip along the sample pad toward the absorbent pad;

[0066] The reagent card is illuminated by a light source, which excites the tracer to produce a second fluorescence;

[0067] Acquire the signal of the second fluorescence during the chromatography reaction, including: the seventh two-dimensional fluorescence signal image I7 of the second fluorescence;

[0068] Acquire the signal of the second fluorescence after the chromatography reaction is completed, including: the eighth two-dimensional fluorescence signal image I8 of the second fluorescence and / or the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence;

[0069] The detection result is output by combining at least a portion of the information of the second fluorescence signal during the chromatography reaction and at least a portion of the information of the second fluorescence signal after the chromatography reaction is completed.

[0070] Optionally, the detection result is output by combining at least some information from I7 and at least some information from I8.

[0071] Optionally, the detection result is output by combining at least some information from I7 and at least some information from C4.

[0072] Optionally, the detection result is output by combining at least some information from I7, at least some information from I8, and at least some information from C4.

[0073] Furthermore, in the immunochromatographic analysis method, the detection line is at least coated with a first internal control; the capture and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the capture;

[0074] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0075] Acquire the signal of the first fluorescence before the detection line during the chromatography reaction, including: the ninth two-dimensional fluorescence signal image I9 of the first fluorescence and / or the fifth one-dimensional fluorescence signal curve C5 of the first fluorescence;

[0076] The detection result is output by combining at least a portion of the information of the second fluorescence signal during the chromatography reaction, at least a portion of the information of the second fluorescence signal after the chromatography reaction is completed, and at least a portion of the information of the first fluorescence signal before the detection line.

[0077] Furthermore, in the immunochromatographic analysis method, the detection line is at least coated with a first internal control; the capture and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the capture;

[0078] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0079] Acquire the signal of the first fluorescence after the chromatography reaction is completed, including: the tenth two-dimensional fluorescence signal image I10 of the first fluorescence and / or the sixth one-dimensional fluorescence signal curve C6 of the first fluorescence;

[0080] The detection result is output by combining at least a portion of the information of the second fluorescence signal during the chromatography reaction, at least a portion of the information of the second fluorescence signal after the chromatography reaction is completed, and at least a portion of the information of the first fluorescence signal before the detection line.

[0081] Furthermore, in the immunochromatographic analysis method, the detection line is at least coated with a first internal control; the capture and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the capture;

[0082] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0083] Acquire the signal of the first fluorescence before the detection line during the chromatography reaction, including: the ninth two-dimensional fluorescence signal image I9 of the first fluorescence and / or the fifth one-dimensional fluorescence signal curve C5 of the first fluorescence;

[0084] Acquire the signal of the first fluorescence after the chromatography reaction is completed, including: the tenth two-dimensional fluorescence signal image I10 of the first fluorescence and / or the sixth one-dimensional fluorescence signal curve C6 of the first fluorescence;

[0085] The detection result is output by combining at least a portion of the information of the second fluorescence signal during the chromatography reaction, at least a portion of the information of the second fluorescence signal after the chromatography reaction is completed, at least a portion of the information of the first fluorescence signal before the detection line during the chromatography reaction, and at least a portion of the information of the first fluorescence signal before the detection line during the chromatography reaction.

[0086] Furthermore, the immunochromatographic analysis method further includes:

[0087] Obtain reference information for the immunochromatographic assay reagent card, the reference information including at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve;

[0088] At least a portion of the reference information is used to output the detection results.

[0089] Furthermore, in the immunochromatographic analysis method described above, obtaining the signal of the second fluorescence during the chromatographic reaction further includes:

[0090] Acquire the signal of the second fluorescence at the first moment during the chromatography reaction, including: the eleventh two-dimensional fluorescence signal image I11 of the second fluorescence at the first moment;

[0091] Acquire the signal of the second fluorescence at the second moment during the chromatography reaction, including: the twelfth two-dimensional fluorescence signal image I12 of the second fluorescence at the second moment;

[0092] At least some information from the fifth two-dimensional fluorescence signal image I11 and at least some information from the sixth two-dimensional fluorescence signal image I12 are used to output the detection result.

[0093] Fifthly, a computer-readable storage medium is provided, the computer-readable storage medium including a program that can be executed by a processor to implement the aforementioned immunochromatographic analysis method.

[0094] Sixthly, an immunochromatographic analyzer is provided, comprising an optical measuring device and a processor. The optical measuring device includes at least an optical imaging unit, which is used to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip. The processor is used to execute a computer-readable storage medium according to this application.

[0095] Furthermore, in the immunochromatographic analyzer, the optical measurement device further includes at least an optical scanning unit, which is used to acquire at least the one-dimensional fluorescence signal curve of the immunochromatographic test strip.

[0096] Furthermore, the optical imaging unit is selected from at least one of CCD and CMOS; the optical scanning unit is a photodetector.

[0097] Furthermore, the immunochromatographic analyzer also includes:

[0098] A sample loading module, for at least the automatic loading and unloading of sample holders or sample tubes; and / or a reagent card loading module, for at least the automatic loading of reagent cards; and / or an incubation module, for at least the carrying and incubation of reagent cards; and / or a temperature control unit, for at least the provision of the temperature conditions required for incubation; and / or a reagent card unloading module, for at least the automatic unloading of reagent cards.

[0099] It should be noted that the immunochromatographic analysis method described in this application is applicable to traditional dry immunochromatographic analysis, semi-liquid immunochromatographic analysis, and whole-liquid immunochromatographic analysis. During the immunochromatographic analysis, a first reagent is provided, comprising a first aptamer conjugated to a tracer. The tracer is used at least to indicate the presence and / or content of the analyte and to generate an optical signal; the optical signal of the tracer is used at least to provide a detection result; simultaneously, the tracer also serves as a second internal control. A first internal control is also coated on the detection line, which is used at least to indicate the amount and / or uniformity of the coating of the analyte captured by the detection line and to generate an optical signal; the optical signals of the tracer and the first internal control are distinguishable from each other. The analyte and the first internal control are independently coated on the detection line; or, the first internal control is directly and / or indirectly bound to the analyte.

[0100] In one embodiment, the immunochromatographic analyzer of this application is used for dry immunochromatographic analysis. The first reagent is located on the test strip, which includes a sample pad, a conjugate pad, an NC membrane, and an absorbent pad arranged sequentially. A detection line is provided on the NC membrane. The first reagent is located on the sample pad and / or the conjugate pad of the test strip. The detection line is coated with a trap, which is at least one of antibody, antigen-binding fragment, aptamer, modified aptamer, aptamer, affinity, antigen, protein, polypeptide, multi-protein complex, exosome, microbial particle / fragment / disc, oligonucleotide, or low molecular weight compound.

[0101] In another embodiment, the immunochromatographic analyzer of this application is used for semi-liquid phase immunochromatographic analysis. The test strip includes a sample pad, an NC membrane, and an absorbent pad arranged sequentially. The NC membrane has a detection line. The first reagent exists independently of the test strip. The detection line is coated with a trap, which is at least one of antibody, antigen-binding fragment, aptamer, modified aptamer, aptamer, affinity, antigen, protein, polypeptide, multi-protein complex, exosome, microbial particle / fragment / disc, oligonucleotide, or low molecular weight compound.

[0102] In another embodiment, the immunochromatographic analyzer of this application is used for total liquid chromatography immunochromatographic analysis. The test strip includes a sample pad, an NC membrane, and an absorbent pad arranged sequentially. The detection line is coated with a trap. The first reagent exists independently of the test strip. The system further includes a second reagent, which is a second aptamer of a coupling linker. The second reagent exists independently of the test strip. The trap is at least one of avidin, streptavidin, anti-biotin antibody, anti-FITC antibody, or anti-DNP antibody.

[0103] Optionally, the first aptamer and the second aptamer specifically recognize and / or bind to the same analyte; or, one of the first aptamer or the second aptamer competes with the analyte to recognize and / or bind to the other aptamer.

[0104] Further, the first aptamer is selected from at least one of antibodies, antigen-binding fragments, aptamers, modified aptamers, aptamers, affinity molecules, antigens, proteins, peptides, multi-protein complexes, exosomes, microbial particles / fragments / debris, oligonucleotides, or low molecular weight compounds. Preferably, the antibody is selected from at least one of monoclonal antibodies, polyclonal antibodies, recombinant antibodies, chimeric antibodies, humanized antibodies, or camel antibodies. Preferably, the antigen-binding fragment is selected from at least one of Fab', Fab, F(ab')2, Fv, and scFV fragments.

[0105] The second aptamer is selected from at least one of antibodies, antigen-binding fragments, aptamers, modified aptamers, aptamers, affinity molecules, antigens, proteins, peptides, multi-protein complexes, exosomes, microbial particles / fragments / debris, oligonucleotides, or low molecular weight compounds. Preferably, the antibody is selected from at least one of monoclonal antibodies, polyclonal antibodies, recombinant antibodies, chimeric antibodies, humanized antibodies, or camel antibodies. Preferably, the antigen-binding fragment is selected from at least one of Fab', Fab, F(ab')2, Fv, and scFV fragments.

[0106] More specifically, when the first aptamer and the second aptamer specifically recognize and / or bind to the same analyte, the first aptamer and the second aptamer can be any molecule capable of specifically recognizing and binding to the analyte.

[0107] It should be understood that the first and second aptamers can be the same or different, as long as they both bind to the analyte, forming a sandwich-like multiplex complex of "tracer-first aptamer-analyte-second aptamer-connector". This complex is captured by the detection zone, and the content or presence of the analyte is calculated from the signal change generated by the tracer. For example, both the first and second aptamers are antibodies; they can be the same antibody or different antibodies. They can be different antibodies binding to the same epitope on the analyte, or they can be different antibodies binding to different epitopes on the analyte. When the first and second aptamers bind to the same epitope of the analyte, the analyte should have two or more repetitions of the same epitope. The first and second aptamers generally bind to different repetitions of the same epitope on the analyte. Furthermore, the signal generated by the tracer in the multiplex complex is proportional to the concentration of the analyte to a certain extent. By measuring the signal generated by the tracer in the detection zone, the presence of the analyte in the sample can be determined, or the concentration of the analyte can be calculated.

[0108] When one of the first aptamers or the second aptamer competes with the analyte to recognize and / or bind to the other aptamer, one of the first aptamers or the second aptamer is any molecule that can compete with the analyte to recognize and / or bind to the other aptamer.

[0109] It should be understood that the first aptamer can compete with the analyte to recognize and / or bind to the second aptamer, or the second aptamer can compete with the analyte to recognize and / or bind to the first aptamer, forming a multiple complex of "tracer-first aptamer-second aptamer-connector" during the competitive reaction with the analyte. The first aptamer and the analyte can be the same or different substances, as long as they can both specifically recognize and / or bind to the second aptamer. By competing with the first aptamer to bind to the second aptamer, and given the known amount of the coupled tracer-first aptamer in the system, the content or presence of the analyte can be calculated from the signal in the detection zone and changes in a preset signal value. Alternatively, the second aptamer and the analyte can be the same or different substances, as long as they can both specifically recognize and / or bind to the first aptamer. By competing with the second aptamer to bind to the first aptamer, and given the known amount of the coupled tracer-first aptamer in the system, the content or presence of the analyte can be calculated from the signal in the detection zone and changes in a preset signal value. Furthermore, by generating signals through tracers in multiple complexes, the signal intensity is inversely proportional to the concentration of the analyte to a certain extent. By measuring the signal generated by the tracers in the detection area, the presence of the analyte in the sample can be determined, or the concentration of the analyte can be calculated.

[0110] The capture and the linker are bound together with certain specificity and affinity. For example, you can choose "biotin-avidin, primary antibody-secondary antibody, antibody-protein, DNP (2,4-dinitrophenyl)-anti-DNP antibody, FITC (fluorescein isothiocyanate)-anti-FITC antibody", etc. Specifically, when the linker is biotin, the capture substance corresponds to avidin and / or streptavidin and / or anti-biotin antibodies; when the linker is avidin and / or streptavidin, the capture substance corresponds to biotin. When the linker is a primary antibody, the capture substance corresponds to a secondary antibody; when the linker is a secondary antibody, the capture substance corresponds to a primary antibody. When the linker is an antibody, the capture substance corresponds to a protein specifically recognized and / or bound by that antibody; when the linker is a protein, the capture substance corresponds to an antibody that can specifically recognize and / or bind to it. When the linker is DNP, the capture substance corresponds to anti-DNP antibodies; when the linker is anti-DNP, the capture substance corresponds to DNP. When the linker is FITC, the capture substance corresponds to anti-FITC antibodies; when the linker is anti-FITC antibodies, the capture substance corresponds to FITC.

[0111] In some specific implementations, the linker and trap connection system is a biotin-streptavidin linker system. It should be understood that one avidin molecule can bind four biotin molecules, and the binding is very stable. Furthermore, avidin and biotin can be conjugated to proteins, enzymes, luciferins, and other molecules without affecting their biological activity. One antibody molecule can conjugate multiple biotin molecules, and avidin can conjugate multiple biotin-antibody conjugates. This multi-stage amplification effect can further improve the sensitivity of the detection system.

[0112] In some specific implementation schemes, the test strip is also covered with a control line, which can be used to determine whether the test is valid or for positioning when the instrument reads the test results.

[0113] The term "specific recognition" or "specific binding" as used in this article can refer to the interaction between an antibody, protein, or peptide and a second chemical substance, wherein the interaction depends on the presence of a specific structure (e.g., an antigenic structure) or a determinant / epitope on the chemical substance, such as an antibody recognizing and binding to a specific antigenic structure, the mutual recognition and binding of a receptor and ligand, or the mutual recognition and binding of biotin and avidin.

[0114] The term "coupled" as used herein broadly refers to a stable bond between two substances resulting from any chemical, physical, or physicochemical interaction (e.g., covalent bond, hydrogen bond, electrostatic interaction, polar attraction, van der Waals attraction, hydrophobic interaction, or adsorption). Unless otherwise stated, this term is intended to encompass both: direct binding / conjugation between two substances, such as the direct binding / conjugation of an antibody to an antigen on a protein, and indirect binding / conjugation between two substances through one or more intermediate means, such as the association between an antibody and a polynucleotide by means of one or more oligonucleotides and / or labels. Therefore, when used with respect to two substances, "coupled" refers to two substances bound / conjugated by any such direct or indirect means.

[0115] The analyte should not be particularly limited and can be any biomolecule or biological cell that needs to be detected and / or quantified in a sample. Non-limiting examples of analytes include antigens, antibodies, antigen-binding fragments, proteins, peptides, multi-protein complexes, hormones, exosomes, oligonucleotides, or low-molecular-weight compounds. The analyte can be detected in any sample of interest and is not particularly limited to biological samples, such as, but not limited to, bodily fluids (e.g., urine, saliva, blood, serum, plasma, sweat), extracts (e.g., cell extracts), and solutions containing proteins and / or DNA (e.g., reaction mixtures).

[0116] The beneficial effects of this application are as follows:

[0117] The inventors of this application discovered that the coating amount of the captured material on the detection line (including the initial coating amount before the chromatographic reaction and the effective coating amount after the chromatographic reaction), the coating uniformity of the captured material on the detection line, and the speed of the chromatographic reaction are key factors affecting the accuracy of the detection results. Therefore, a light source assembly is set up to excite a first internal reference to generate first fluorescence, and an optical measurement device including an optical imaging unit and an optical scanning unit is set up to acquire the fluorescence signal. Specifically, the optical imaging unit is used to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip, and the optical scanning unit is used to acquire a one-dimensional fluorescence signal curve of the immunochromatographic test strip. Then, the detection results are corrected using at least some information from the two-dimensional fluorescence signal image and / or at least some information from the one-dimensional fluorescence signal curve to obtain corrected detection results, which can effectively ensure the accuracy of the detection results.

[0118] Meanwhile, by combining the two-dimensional fluorescence signal image of the second internal reference, the chromatography process is dynamically monitored, enabling high-precision detection of analytes; reducing the difference between raw material rolls of the NC membrane, the complexity of production is reduced; and the requirements for the detection environment, such as temperature and humidity, are reduced.

[0119] Furthermore, the immunochromatographic analysis method of this application is applicable to traditional dry immunochromatographic analysis, semi-liquid immunochromatographic analysis, and whole-liquid immunochromatographic analysis, and has a wide range of applicable scenarios, which can effectively improve the detection accuracy of various immunochromatographic modes. Attached Figure Description

[0120] Figure 1 is a schematic diagram of one embodiment of the immunochromatographic analyzer of the present invention.

[0121] Figure 2 is a schematic diagram of another embodiment of the immunochromatographic analyzer of the present invention.

[0122] Figure 3 is a two-dimensional fluorescence signal image of the first fluorescence before the detection line in one case of chromatography reaction.

[0123] Figure 4 shows a two-dimensional fluorescence signal image of the first fluorescence before the detection line in another case.

[0124] Figure 5 shows a one-dimensional fluorescence signal curve of the first fluorescence before the detection line in one scenario.

[0125] Figure 6 shows the signal of the first fluorescence after the chromatography reaction is completed in one scenario.

[0126] Figure 6(a) is a two-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction is completed in one case.

[0127] Figure 6(b) is a one-dimensional fluorescence signal curve of the first fluorescence after the chromatography reaction is completed in one case.

[0128] Figure 6(c) is a three-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction is completed in one case.

[0129] Figure 7 shows two-dimensional fluorescence signal images of the second fluorescence at two different times during chromatography in one scenario. Detailed Implementation

[0130] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0131] As shown in Figure 1, an immunochromatographic analyzer 1 includes a sample acquisition device 11, a reagent supply device 12, a sample transfer device 13, a light source assembly 14, an optical measurement device 15, and a data processing unit 17.

[0132] The sample collection device 11 is used to collect samples and transport them to a sample reaction device (not shown) for sample preparation. In some embodiments, the sample collection device 11 includes a sampling needle, a syringe, and a sampling needle cleaning swab (not shown). Of course, the sample collection device 11 is not limited to this form and can be configured according to needs. For example, in some other embodiments, the sample collection device 11 may also include a metering pump.

[0133] The reagent supply device 12 is used to provide reagents that can react with the sample and participate in the sample preparation process. In some embodiments, the reagents include at least a diluent and a lysing agent. In other embodiments, the reagents include liquid reagents and solid reagents. In still other embodiments, at least a portion of the reagents is provided separately. For example, the diluent is provided separately, contained in a diluent bottle, and the diluent contained in the diluent bottle is sufficient for multiple tests (e.g., 50 tests); the lysing agent is provided separately, contained in a lysing agent bottle, and the lysing agent contained in the lysing agent bottle is sufficient for multiple tests (e.g., 30 tests). In still other embodiments, at least a portion of the reagents is provided via a reagent card. For example, the reagent may be a first reagent disposed on the conjugate pad of a test strip on the reagent card; or, for instance, the reagent may be a lyophilized reagent disposed in a cavity in a non-test strip area of ​​the reagent card.

[0134] The sample transfer device 13 is configured to transfer the sample to the test strip of the immunochromatographic analysis reagent card. The light source assembly 14 is used to excite a first fluorescent substance to produce first fluorescence and to excite a second fluorescent substance to produce second fluorescence. The optical measurement device 15 includes an optical imaging unit and an optical scanning unit (not shown in Figure 1). The optical imaging unit is used to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip, and the optical scanning unit is used to acquire a one-dimensional fluorescence signal curve of the immunochromatographic test strip. Optionally, the sample acquisition device 11 and the sample transfer device 13 can be different devices or the same device.

[0135] In one embodiment, the light source assembly includes a light source capable of simultaneously emitting at least two different wavelengths of excitation light: a first excitation light of a first wavelength and a second excitation light of a second wavelength. The first excitation light is used to excite a first fluorescent substance to produce first fluorescence, and the second excitation light is used to excite a second fluorescent substance to produce second fluorescence. In another embodiment, the light source assembly includes two light sources, which respectively emit the first excitation light of a first wavelength and the second excitation light of a second wavelength. In still some embodiments, the light source assembly includes a light source emitting a third excitation light of a third wavelength, which is capable of exciting both the first fluorescent substance to produce first fluorescence and the second fluorescent substance to produce second fluorescence.

[0136] In one embodiment, the optical imaging unit includes a CCD. In some other embodiments, the optical imaging unit includes a CMOS sensor. In one embodiment, the optical scanning unit includes a photodetector.

[0137] In this embodiment, the optical measuring device further includes a first filter and a second filter, wherein the first filter is used to filter out the first fluorescence and the second filter is used to filter out the second fluorescence.

[0138] In some alternative implementations, as shown in FIG2, the immunochromatographic analyzer 1 further includes a control unit 16.

[0139] In this embodiment, the test strip of the immunochromatographic analysis reagent card includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with a trap and a first fluorescent substance, which serves as a first internal control. The fluorescent signal substance can be at least one of fluorescent microspheres, quantum dots, fluorescent proteins, or luciferin molecules. The sample includes a tracer, which is used to indicate the content and / or presence of the analyte and also serves as a second internal control. Before the chromatographic reaction, the second internal control is present in the sample. During the chromatographic reaction, the second internal control precipitates along the chromatographic direction of the test strip towards the detection line. In some optional embodiments, the second internal control may also be present in the non-detection line region of the test strip.

[0140] In some alternative embodiments, the trap and the first intrinsic parameter are independently included in the detection line. In other alternative embodiments, the first intrinsic parameter is directly and / or indirectly incorporated into the trap.

[0141] In this embodiment, the first internal parameter is directly bound to the captured object, and the coating amount of the first internal parameter is highly consistent with the coating amount of the captured object; the coating uniformity of the first internal parameter is also highly consistent with the coating uniformity of the captured object.

[0142] In an optional embodiment, the control unit 16 controls the optical measuring device 15 to acquire the first one-dimensional fluorescence signal curve C1 of the first fluorescence before the detection line in the chromatography reaction. From the first one-dimensional fluorescence signal curve C1, information such as the peak intensity and / or peak area of ​​the fluorescence signal corresponding to the detection line can be obtained, and the actual coating amount of the captured substance on the detection line of the test strip can be obtained.

[0143] In a preferred embodiment, the control unit 16 controls the optical measuring device 15 to acquire the first two-dimensional fluorescence signal curve I1 of the first fluorescence before the detection line in the chromatography reaction. From the first two-dimensional fluorescence signal curve I1, information such as the intensity and intensity distribution of the fluorescence signal corresponding to the detection line can be obtained, and the actual coating uniformity of the captured material on the detection line of the test strip can be obtained.

[0144] The actual coating amount and / or uniformity of the captured material on the detection line can affect the detection results. Therefore, it is necessary to obtain the first one-dimensional fluorescence signal curve C1 and / or the first two-dimensional fluorescence signal curve I1 from the start of the chromatographic reaction to the detection line, and correct the detection results based on at least some of the information therein to obtain the corrected detection results. Specifically, at least one detection result correction parameter can be obtained first based on at least some of the information, and then the detection results can be corrected using the at least one detection result correction parameter.

[0145] Figures 3 and 4 show two scenarios of two-dimensional fluorescence signal images of the first fluorescence acquired by the optical imaging unit before the chromatographic reaction proceeds to the detection line (specifically, before sample addition). In Figure 3, the test strip includes a detection line (T line) and a control line (C line). Both the detection line and the control line are coated with the first fluorescent substance. In Figure 4, the test strip includes a first detection line (T1 line), a second detection line (T2 line), and a control line (C line). The first detection line (T1 line), the second detection line (T2 line), and the control line (C line) are all coated with the first fluorescent substance. The two-dimensional fluorescence signal images shown in Figures 3 and 4 can provide more information about the fluorescence signal, including at least information such as fluorescence signal intensity, fluorescence signal uniformity, and signal location.

[0146] Figure 5 shows one scenario of a one-dimensional fluorescence signal image of the first fluorescence acquired by the optical scanning unit before the detection line during the chromatography reaction. In Figure 5, the test strip includes a detection line (T line) and a control line (C line). The one-dimensional fluorescence signal image shown in Figure 5 provides relatively little fluorescence signal information, generally including information such as fluorescence signal intensity and signal location, but not information on fluorescence signal uniformity.

[0147] In another embodiment, the control unit 16 controls the optical measurement device 15 to acquire the second one-dimensional fluorescence signal curve C2 and the second two-dimensional fluorescence signal curve I2 of the first fluorescence after the chromatographic reaction is completed. This allows the effective coating amount and uniformity of the captured material on the detection line of the test strip after the chromatographic reaction. Since the captured material may be lost during the chromatographic reaction, the actual coating amount and actual coating uniformity of the captured material on the detection line before the chromatographic reaction reaches the detection line are not equal to the effective coating amount and effective coating uniformity of the captured material on the detection line. Therefore, it is necessary to acquire the second one-dimensional fluorescence signal curve C2 and / or the second two-dimensional fluorescence signal curve I2 of the first fluorescence after the chromatographic reaction is completed, and correct the detection results based on at least some of the information therein, to obtain the corrected detection results.

[0148] Figure 6 shows the first fluorescence signal after the chromatography reaction is completed. Figure 6(a) is a two-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction, acquired by the optical imaging unit; Figure 6(b) is a one-dimensional fluorescence signal curve of the first fluorescence of the same test strip after the chromatography reaction, acquired by the optical scanning unit; Figure 6(c) is a three-dimensional fluorescence signal image converted from Figure 6(a). The two-dimensional fluorescence signal image in Figure 6(a) includes information such as fluorescence signal intensity, fluorescence signal uniformity, and signal location. The one-dimensional fluorescence signal image in Figure 6(b) only includes fluorescence signal intensity and signal location information. The three-dimensional fluorescence signal image in Figure 6(c) clearly demonstrates the uniformity information of the fluorescence signal.

[0149] In one specific embodiment, the data processing unit 17 first obtains at least one detection result correction parameter based on at least a portion of the information from the first one-dimensional fluorescence signal curve C1, and / or the first two-dimensional fluorescence signal curve I1, and / or the second one-dimensional fluorescence signal curve C2, and / or the second two-dimensional fluorescence signal curve I2, and then corrects the detection result using the obtained at least one detection result correction parameter. The specific detection result correction parameter can be 1, 2, 3, 4, 5, etc., and correspondingly, the obtained corrected detection result can also be 1, 2, 3, 4, 5, etc.

[0150] Due to the loss of the captured material, the tracer material it captured is also carried away, resulting in a lower amount of tracer material captured in the detection line area than expected, leading to inaccurate detection results. Therefore, in some preferred embodiments, the data processing unit 17 combines at least one of C1 and I1 and at least one of C2 and I2 to obtain information on the loss of the captured material during the chromatography process on the detection line (e.g., loss amount, loss rate, loss uniformity, etc.), and corrects the detection results to obtain corrected detection results. Specifically, at least one detection result correction parameter can be obtained first, and then the detection result can be corrected using the obtained at least one detection result correction parameter. More specifically, the detection result correction parameters can be 1, 2, 3, 4, 5, etc., and correspondingly, the obtained corrected detection results can also be 1, 2, 3, 4, 5, etc.

[0151] In a preferred embodiment, the control unit 16 controls the optical measurement device 15 to acquire a third two-dimensional fluorescence signal image I3 of the second fluorescence during the chromatography reaction.

[0152] In a more preferred embodiment, the control unit 16 controls the optical measurement device 15 to acquire a fourth two-dimensional fluorescence signal image I4 and a fifth two-dimensional fluorescence signal image I5 of the second fluorescence at two different times during the chromatography reaction. The two different times are time 1 (denoted as t1) and time 2 (denoted as t2). Figure 7 shows one scenario of two-dimensional fluorescence signal images of the second fluorescence at two different times during the chromatography process. After simple data processing, one-dimensional fluorescence signal curves of the two-dimensional fluorescence signal images at the two times can be obtained, thereby obtaining the corresponding chromatography position. Alternatively, the position of the chromatography reaction at the current time can be determined directly based on the position information of the fluorescence signal contained in the two-dimensional fluorescence signal image, and then the rate of the chromatography reaction can be obtained using the position difference and time difference between the two different times. For example, chromatography rate = (chromatographic position 2 - chromatographic position 1) / (time 2 - time 1).

[0153] The data processing unit 17 can obtain information related to the rate of the chromatographic reaction based on the third two-dimensional fluorescence signal image I3. Since the amount of tracer captured by the trap will differ depending on the chromatographic reaction rate, this application configures the data processing unit to correct the detection results using at least some information from the third two-dimensional fluorescence signal image I3, obtaining corrected detection results and improving the accuracy of the detection results.

[0154] Preferably, the data processing unit 17 can also obtain information related to the rate of the chromatographic reaction based on the fourth two-dimensional fluorescence signal image I4 and the fifth two-dimensional fluorescence signal image I5. Based on the current position of the chromatographic reaction in the two-dimensional fluorescence signal images at two different times, information related to the rate of the chromatographic reaction can be better reflected. Accordingly, this application sets the data processing unit to correct the detection results using at least some information from the fourth two-dimensional fluorescence signal image I4 and the fifth two-dimensional fluorescence signal image I5, obtaining corrected detection results and improving the accuracy of the detection results.

[0155] In a preferred embodiment, the immunochromatographic analyzer 1 further includes an information acquisition unit (not shown in FIG. 1). The information acquisition unit can identify the label information of the reagent card and acquire the reference information of the current reagent card. Generally, the reference information of the reagent card includes, but is not limited to, at least one of the following: the reference concentration information R0 of the first fluorescent substance on the detection line of this batch of reagent cards, the reference fluorescence signal T0 of the first fluorescent substance on the detection line of this batch of reagent cards, the reference chromatography speed V0 of the test strip of this batch of reagent cards, the analyte information of this reagent card, and the calibration curve of this batch of reagent cards.

[0156] Accordingly, in a preferred embodiment, the data processing unit 17 combines at least some of the information in these reference information, at least some of the information in the two-dimensional fluorescence signal image and / or at least some of the information in the one-dimensional fluorescence signal curve to correct the detection result, thereby obtaining the corrected detection result.

[0157] Specifically, the data processing unit 17 combines at least some of the information from these reference information, as well as at least some of the information from the two-dimensional fluorescence signal image and / or at least some of the information from the one-dimensional fluorescence signal curve to first obtain at least one detection result correction parameter, and then uses the obtained at least one detection result correction parameter to correct the detection result. More specifically, the detection result correction parameter can be 1, 2, 3, 4, 5, etc., and correspondingly, the obtained corrected detection result can also be 1, 2, 3, 4, 5, etc.

[0158] In this embodiment, the correction parameters consist of at least one correction factor. The correction factors include at least: a first capture coating amount correction factor a, related to the amount of capture on the detection line before the chromatographic reaction proceeds to the detection line; a second capture coating amount correction factor b, related to the amount of capture on the detection line after the chromatographic reaction is completed; a first capture coating uniformity correction factor c, related to the amount of capture on the detection line before the chromatographic reaction proceeds to the detection line; a second capture coating uniformity correction factor d, related to the amount of capture on the detection line after the chromatographic reaction is completed; and an actual chromatography rate correction factor e, related to the actual chromatographic reaction rate. The completion of the chromatographic reaction generally refers to a specific time point after sample addition. In this embodiment, the completion of the chromatographic reaction refers to the 5th minute after sample addition. Accordingly, the correction parameters include at least one of a, b, c, d, and e, any combination of two, any combination of three, any combination of four, or a combination of five.

[0159] In some other alternative embodiments, when the data processing unit 17 obtains correction parameters or correction factors through data processing, it may combine at least a portion of information such as the background signal of the reagent card and / or test strip, the HCT value of the sample, and the dilution ratio of the sample for data processing.

[0160] The chromatographic reaction in this application includes physical and / or chemical and / or biological reactions of the liquid on the test strip. The completion of the chromatographic reaction generally refers to a specific time point after sample addition; for example, the completion of the chromatographic reaction can be any time point between 30 seconds and 15 minutes after sample addition.

[0161] The following is an example, using the combination of modified parameters (a, b).

[0162] After the chromatography reaction is completed (in this example, the time point after sample addition is taken as the time point after the chromatography reaction is completed), a one-dimensional fluorescence signal curve of the tracer (second fluorescent substance, also used as second internal reference) is obtained. The detection result X of the analyte is obtained based on the peak intensity in the one-dimensional fluorescence signal curve.

[0163] Two-dimensional fluorescence signal images of the first fluorescence generated by the first internal control before sample addition and after the chromatography reaction are acquired respectively. The detection results are corrected based on the ratio of the average fluorescence intensity in the detection line region in the two two-dimensional fluorescence signal images (for example, the average fluorescence intensity in the detection line region in the two-dimensional fluorescence signal image of the first fluorescence before sample addition is a, and the average fluorescence intensity in the detection line region in the two-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction is b, and the ratio is n = b / a, where n represents the effective coating rate of the first internal control before and after the chromatography reaction).

[0164] As an example, the method for correcting the detection results using correction parameters is as follows: the corrected detection result is Y = f(a, b, X) = K * X * a / b. Where K is the temperature compensation factor.

[0165] In one alternative implementation, either a or b can be used as the correction parameter. The reference fluorescence signal T0 of the first fluorescent substance on the detection line of the reagent card in this batch is obtained, and the intensity of the reference fluorescence signal T0 is denoted as a0. As an example, a method for correcting the detection result using the correction parameter is: the corrected detection result is Y = f(a, X) = X * a0 / a, or Y = f(b, X) = X * a0 / b.

[0166] In another alternative implementation, the two-dimensional fluorescence signal image of the first fluorescence generated by the first intrinsic parameter can be replaced by the one-dimensional fluorescence signal curve of the first fluorescence generated by the first intrinsic parameter. Accordingly, the peak intensity, peak area, etc. in the one-dimensional fluorescence signal curve can be used as the characteristic information of the first fluorescence for calculation.

[0167] A preferred embodiment involves acquiring a two-dimensional fluorescence signal image of the first fluorescence generated by the first internal reference. The two-dimensional fluorescence signal image also includes information related to "uniformity".

[0168] Taking the combination of correction parameters (a, b, c, d) as an example, the uniformity of the first fluorescence before sample addition is taken as c (for example, the detection line region in the two-dimensional fluorescence signal image of the first fluorescence before sample addition can be divided into 100 small rectangular regions of the same size, the fluorescence signal intensity value in each small rectangular region can be obtained, and then the uniform distribution standard deviation of these fluorescence signal intensity values ​​can be calculated, and the uniform distribution standard deviation can be taken as uniformity c). The uniformity of the first fluorescence after the chromatography reaction is taken as d (for example, the detection line region in the two-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction is divided into 100 small rectangular regions of the same size, the fluorescence signal intensity value in each small rectangular region can be obtained, and then the uniform distribution standard deviation of these fluorescence signal intensity values ​​can be calculated, and the uniform distribution standard deviation can be taken as uniformity d).

[0169] As an example, the method of correcting the detection result with correction parameters is: the corrected detection result is Y = f(a, b, c, d, X) = X * a * a(bd) / [b * b * (ac)].

[0170] In a more preferred embodiment, information related to the chromatography reaction rate is also included. As an example, the following describes a combination of correction parameters (a, b, c, d, e).

[0171] Two-dimensional fluorescence signal images of the second internal control (i.e., the tracer in this embodiment) at 10 s and 15 s during incubation were obtained during the chromatography reaction. The rate of the chromatography reaction was obtained by combining the two two-dimensional fluorescence signal images (for example, if the time difference between the two two-dimensional fluorescence signal images is 5 s, comparing the two two-dimensional fluorescence signals, the positional difference of the fluorescence signal can be obtained as D, and the rate of the chromatography reaction can be obtained by dividing the positional difference D by the time difference). This rate can be used as a correction factor e.

[0172] As an example, the method of correcting the detection result with correction parameters is as follows: The corrected detection result is Y = f(a, b, c, d, e, X) = X * a * a(bd) / [b * b * (ac) * e].

[0173] In some optional embodiments, the completion of the chromatographic reaction generally refers to a specific time point after sample addition. For example, the completion of the chromatographic reaction can be any time point between the 30th second and the 15th minute after sample addition. In a preferred embodiment, the time interval between acquiring the light signal T of the tracer and acquiring the second light signal T2 of the first internal reference is less than or equal to 5 minutes. For example, it can be 4 minutes, 3 minutes, 2 minutes, 1 minute, 30 seconds, 20 seconds, 10 seconds, 5 seconds, or 0 seconds.

[0174] In some alternative implementations, as shown in FIG2, the immunochromatographic analyzer 1 further includes a result output unit 18.

[0175] In a preferred embodiment, the result output unit 18 includes a display interface for displaying the detection result and a corrected detection result. In another preferred embodiment, the result output unit 18 includes a display interface for simultaneously displaying the detection result, a corrected detection result, and the correction parameter or correction factor corresponding to the corrected detection result.

[0176] In some optional embodiments, the result output unit 18 includes a display interface that simultaneously displays the detection result and three different corrected detection results, along with their corresponding correction parameters or correction factors. Furthermore, the detection result or corrected detection result displayed on the display interface can be selected. Preferably, the selected detection result or corrected detection result is taken as the final detection result.

[0177] In some preferred embodiments, the result output unit 18 includes a display interface that displays at least one of a two-dimensional fluorescence signal image and a three-dimensional fluorescence signal image of the first fluorescence and / or the second fluorescence.

[0178] In some alternative embodiments, the immunochromatographic analyzer further includes: a sample loading module for automatically loading and unloading sample holders or sample tubes; and / or a reagent card loading module for automatically loading reagent cards; and / or an incubation module for carrying and incubating reagent cards; and / or a temperature control unit for providing the temperature conditions required for incubation; and / or a reagent card unloading module for automatically unloading reagent cards.

[0179] Another embodiment of this application relates to an immunochromatographic analysis method, comprising:

[0180] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. The immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least a first internal control and a trap. The trap and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the trap.

[0181] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0182] The sample is fed to the test strip of the immunochromatographic assay reagent card to initiate the chromatographic reaction; the sample contains a tracer;

[0183] The reagent card is illuminated by a light source, which excites the tracer to produce a second fluorescence;

[0184] Acquire the signal of the first fluorescence; the signal of the first fluorescence includes: a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the immunochromatographic analysis reagent card is provided to the immunochromatographic analyzer and the chromatography reaction proceeds to the detection line; and a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the chromatography reaction is completed;

[0185] Acquire the signal of the second fluorescence; the signal of the second fluorescence includes: a two-dimensional fluorescence signal image of the second fluorescence and / or a one-dimensional fluorescence signal curve of the second fluorescence after the chromatography reaction is completed;

[0186] The detection result is output by combining at least a portion of the information from the first fluorescence signal and at least a portion of the information from the second fluorescence signal.

[0187] In a preferred embodiment, the immunochromatographic analysis method further includes:

[0188] Obtain reference information for the immunochromatographic assay reagent card, the reference information including at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve;

[0189] The detection result is output by combining at least a portion of the information from the first fluorescence signal, at least a portion of the information from the second fluorescence signal, and at least a portion of the information from the reference information.

[0190] Another embodiment of this application relates to an immunochromatographic analysis method, comprising:

[0191] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. The immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least a first internal control and a trap. The trap and the first internal control are independently coated on the detection line, or the first internal control is directly and / or indirectly bound to the trap.

[0192] The light source illuminates the reagent card and excites the first internal control to produce a first fluorescence;

[0193] The sample is fed to the test strip of the immunochromatographic assay reagent card to initiate the chromatographic reaction; the sample contains a tracer;

[0194] The reagent card is illuminated by a light source, which excites the tracer to produce a second fluorescence;

[0195] Acquire the signal of the first fluorescence; the signal of the first fluorescence includes: a two-dimensional fluorescence signal image and / or a one-dimensional fluorescence signal curve of the first fluorescence after the immunochromatographic analysis reagent card is provided to the immunochromatographic analyzer and the chromatography reaction proceeds to the detection line and / or after the chromatography reaction is completed;

[0196] Acquire the signal of the second fluorescence; the signal of the second fluorescence includes: a two-dimensional fluorescence signal image of the second fluorescence and / or a one-dimensional fluorescence signal curve of the second fluorescence after the chromatography reaction is completed, and a two-dimensional fluorescence signal image of the second fluorescence during the chromatography reaction.

[0197] The detection result is output by combining at least a portion of the information from the first fluorescence signal and at least a portion of the information from the second fluorescence signal.

[0198] In a preferred embodiment, the immunochromatographic analysis method further includes:

[0199] Obtain reference information for the immunochromatographic assay reagent card, the reference information including at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve;

[0200] The detection result is output by combining at least a portion of the information from the first fluorescence signal, at least a portion of the information from the second fluorescence signal, and at least a portion of the information from the reference information.

[0201] In a first embodiment, an immunochromatographic analysis method is provided, comprising:

[0202] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. In this embodiment, the immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least one trap. The trap is used to capture tracers during the chromatography process. In this embodiment, the trap is an antibody. In some other optional embodiments, the trap can be at least one of the following: antigen-binding fragments, aptamers, modified aptamers, aptamers, affinity molecules, antigens, proteins, peptides, multi-protein complexes, exosomes, biotin, FITC, DNP, microbial particles / fragments / debris, oligonucleotides, or low molecular weight compounds.

[0203] A sample containing a tracer is provided to a reagent card, such that the tracer in the sample is chromatographically deposited on the test strip along the sample pad towards the absorbent pad. In this embodiment, the tracer is a fluorescent signaling substance, specifically fluorescent microspheres. In some alternative embodiments, the fluorescent signaling substance may be at least one of fluorescent microspheres, quantum dots, fluorescent proteins, or luciferin molecules.

[0204] The light source illuminates the reagent card, exciting the tracer to produce a second fluorescence.

[0205] Acquire the signal of the second fluorescence during the chromatography reaction, including: the seventh two-dimensional fluorescence signal image I7 of the second fluorescence.

[0206] Acquiring the signal of the second fluorescence after the chromatography reaction is completed includes: an eighth two-dimensional fluorescence signal image I8 of the second fluorescence. In some optional embodiments, a fourth one-dimensional fluorescence signal curve C4 of the second fluorescence after the chromatography reaction is completed may also be acquired, or both the eighth two-dimensional fluorescence signal image I8 and the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence may be acquired simultaneously.

[0207] Finally, the detection results are combined with those from I7 and I8. In some optional implementations, the detection results can also be combined with those from I7 and C4, or they can be combined with those from I7, I8, and C4.

[0208] As an example, a correction factor related to the chromatography rate of the test strip used in this embodiment can be obtained from I7 (e.g., by combining the sample addition time point, the acquisition time point of I7, and the chromatography reaction position in I7, the chromatography reaction rate can be obtained, and thus a correction factor related to the chromatography rate can be obtained), denoted as e. The amount of captured tracer can be obtained from I8, thus obtaining an initial detection result, denoted as X. Then, the initial detection result X is corrected using the chromatography rate-related correction factor e to obtain the final detection result, denoted as Y1. For example, the final detection result Y1 = f(e, X) = X / e. The final detection result Y1 is then output.

[0209] As another example, the uniformity of the captured tracer can be obtained from I8, and a correction factor f related to the tracer uniformity can be obtained. Then, the initial detection result X is corrected using the correction factor e related to the chromatography rate and the correction factor f related to the tracer uniformity to obtain the final detection result, denoted as Y2. For example, the final detection result Y2 = f(e, X) = X*f / e. The final detection result Y2 is then output.

[0210] In a second embodiment, an immunochromatographic analysis method is provided, comprising:

[0211] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. In this embodiment, the immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with at least one trap. The trap is used to capture tracers during the chromatography process. In this embodiment, the trap is an antigen. In some other optional embodiments, the trap can be at least one of the following: antigen-binding fragments, aptamers, modified aptamers, aptamers, affinity molecules, antibodies, proteins, peptides, multi-protein complexes, exosomes, biotin, FITC, DNP, microbial particles / fragments / debris, oligonucleotides, or low molecular weight compounds.

[0212] A sample containing a tracer is provided to a reagent card, such that the tracer in the sample is chromatographically deposited on the test strip along the sample pad towards the absorbent pad. In this embodiment, the tracer is a fluorescent signaling substance, specifically fluorescent microspheres. In some alternative embodiments, the fluorescent signaling substance may be at least one of fluorescent microspheres, quantum dots, fluorescent proteins, or luciferin molecules.

[0213] The light source illuminates the reagent card, exciting the tracer to produce a second fluorescence.

[0214] The eleventh two-dimensional fluorescence signal image I11 of the second fluorescence at the first moment during the chromatography reaction was acquired; the twelfth two-dimensional fluorescence signal image I12 of the second fluorescence at the second moment during the chromatography reaction was acquired.

[0215] Acquiring the signal of the second fluorescence after the chromatography reaction is completed includes: the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence. In some optional embodiments, an eighth two-dimensional fluorescence signal image I8 of the second fluorescence after the chromatography reaction is completed may also be acquired, or the eighth two-dimensional fluorescence signal image I8 of the second fluorescence and the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence may be acquired simultaneously.

[0216] Finally, the detection results are output by combining C4, I11, and I12. In some optional implementations, the detection results can also be output by combining I8, I11, and I12, or by combining I8, C4, I11, and I12.

[0217] In this context, "completion of chromatographic reaction" generally refers to a specific time point after sample addition. For example, "completion of chromatographic reaction" can be any time point between 30 seconds and 15 minutes after sample addition.

[0218] As an example, correction factors related to the chromatography speed of the test strip used in this embodiment can be obtained from I11 and I12.

[0219] For example, Figure 7 shows a scenario where two-dimensional fluorescence signal images of the second fluorescence at two different moments during chromatography (corresponding to I11 and I12, respectively). After simple data processing, one-dimensional fluorescence signal curves of the two-dimensional fluorescence signal images at the two moments can be obtained, thus revealing the corresponding chromatography positions. Alternatively, the position of the chromatography reaction at the current moment can be determined directly based on the positional information of the fluorescence signal contained in the two-dimensional fluorescence signal image. Then, the rate of the chromatography reaction can be obtained using the position difference and time difference between the two different moments. For example, chromatography rate = (chromatographic position 2 - chromatography position 1) / (time 2 - time 1). Furthermore, a correction factor related to the chromatography rate, denoted as e, can be obtained. The amount of tracer captured from C4 is obtained, resulting in an initial detection result, denoted as X. Then, the initial detection result X is corrected using the chromatography rate-related correction factor e to obtain the final detection result, denoted as Y3. For example, the final detection result Y3 = f(e, X) = m*X / e, where m is a correction coefficient constant. Output the final detection result Y3.

[0220] In a third embodiment, an immunochromatographic analysis method is provided, comprising:

[0221] An immunochromatographic analysis reagent card is provided to an immunochromatographic analyzer. In this embodiment, the immunochromatographic analysis reagent card includes a test strip, which includes at least a sample pad, an NC membrane, and an absorbent pad. The sample pad is located upstream of the NC membrane, and a detection line is provided on the NC membrane. The detection line is coated with a trap and a first internal control. The trap is used to capture tracers during the chromatography process. In this embodiment, the trap is a direct conjugate of biotin and the first internal control, which is coated together on the detection line. In some other optional embodiments, the trap and the first internal control may be independently coated on the detection line, or the first internal control may be indirectly bound to the trap.

[0222] The captured substance can also be at least one of the following: antigen-binding fragment, aptamer, modified aptamer, aptamer, affinity compound, antibody, protein, peptide, multiprotein complex, exosome, antigen, FITC, DNP, microbial particle / fragment / disc, oligonucleotide, or low molecular weight compound. The first internal control is a fluorescent signal substance, which is different from the tracer.

[0223] A sample containing a tracer is provided to a reagent card, such that the tracer in the sample is chromatographically deposited on the test strip along the sample pad towards the absorbent pad. In this embodiment, the tracer is a fluorescent signaling substance, specifically fluorescent microspheres. In some alternative embodiments, the fluorescent signaling substance may be at least one of fluorescent microspheres, quantum dots, fluorescent proteins, or luciferin molecules.

[0224] The light source illuminates the reagent card, exciting the first internal control to produce the first fluorescence and exciting the tracer to produce the second fluorescence.

[0225] A ninth two-dimensional fluorescence signal image I9 of the first fluorescence is acquired before the detection line during the chromatography reaction. In some optional embodiments, a fifth one-dimensional fluorescence signal curve C5 of the first fluorescence can be acquired; or, both the ninth two-dimensional fluorescence signal image I9 and the fifth one-dimensional fluorescence signal curve C5 of the first fluorescence can be acquired simultaneously. In other optional embodiments, a tenth two-dimensional fluorescence signal image I10 of the first fluorescence and / or a sixth one-dimensional fluorescence signal curve C6 of the first fluorescence are also acquired after the chromatography reaction is completed (in this example, the time point after sample addition is taken as the completion time of the chromatography reaction).

[0226] The eleventh two-dimensional fluorescence signal image I11 of the second fluorescence at the first moment during the chromatography reaction was acquired; the twelfth two-dimensional fluorescence signal image I12 of the second fluorescence at the second moment during the chromatography reaction was acquired.

[0227] Acquiring the signal of the second fluorescence after the chromatography reaction is completed includes: the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence. In some optional embodiments, an eighth two-dimensional fluorescence signal image I8 of the second fluorescence after the chromatography reaction is completed may also be acquired, or the eighth two-dimensional fluorescence signal image I8 of the second fluorescence and the fourth one-dimensional fluorescence signal curve C4 of the second fluorescence may be acquired simultaneously.

[0228] Finally, the detection results are output by combining I9, C4, I11, and I12. In some optional implementations, the detection results can also be output by combining C5, I8, I11, and I12; or by combining I9, C5, I8, C4, I11, and I12; or by combining I10, C5, I9, I11, and I12; or by combining C5, C6, I8, I11, and I12; or by combining I9, I10, I8, I11, and I12. Other scenarios are not listed here.

[0229] As an example, correction factors related to the amount of trap coating and / or coating uniformity on the detection line before and after sample addition can be obtained from I9 and I10, respectively.

[0230] For example, Figure 3 shows a scenario where the optical imaging unit acquires a two-dimensional fluorescence signal image of the first fluorescence before the chromatographic reaction proceeds to the detection line (specifically, before sample application). In Figure 3, the test strip includes a detection line (T line) and a control line (C line). Both the detection line and the control line are coated with the first fluorescent substance. The two-dimensional fluorescence signal image shown in Figure 3 can provide more information about the fluorescence signal, including at least the fluorescence signal intensity, fluorescence signal uniformity, and signal location. With simple data processing, information related to the amount of trap coating on the detection line before sample application and / or the coating uniformity can be obtained from Figure 3.

[0231] Figure 6 shows one scenario of a two-dimensional fluorescence signal image of the first fluorescence acquired by the optical imaging unit after the chromatography reaction. Figure 6(a) is the two-dimensional fluorescence signal image of the first fluorescence acquired by the optical imaging unit after the chromatography reaction; Figure 6(b) is the one-dimensional fluorescence signal curve of the first fluorescence of the same test strip after the chromatography reaction, acquired by the optical scanning unit; Figure 6(c) is the three-dimensional fluorescence signal image converted from Figure 6(a). The two-dimensional fluorescence signal image in Figure 6(a) includes information such as fluorescence signal intensity, fluorescence signal uniformity, and signal location. The one-dimensional fluorescence signal image in Figure 6(b) only includes fluorescence signal intensity and signal location information. The three-dimensional fluorescence signal image in Figure 6(c) clearly demonstrates the uniformity information of the fluorescence signal.

[0232] The average fluorescence intensity within the detection line region of the first fluorescence signal image before sample addition can be obtained from I9 and I10 (or C5 and C6), denoted as 'a'. The average fluorescence intensity within the detection line region of the first fluorescence signal image after the chromatography reaction is completed is denoted as 'b'. The ratio is n = b / a, where n represents the effective coating rate of the first internal reference before and after the chromatography reaction. Then, this ratio (i.e., the amount of capture material before and after the chromatography reaction) and a correction factor e related to the chromatography rate are used to correct the initial detection result X, yielding the final detection result, denoted as Y4. For example, the final detection result Y4 = f(a, b, e, X) = K*X*a / (b*e), where K is the temperature compensation factor.

[0233] As another example, the uniformity of the first fluorescence before sample addition can be obtained from I9 and I10, denoted as c (for example, the detection line region in the two-dimensional fluorescence signal image of the first fluorescence before sample addition can be divided into 100 small rectangular regions of the same size, the fluorescence signal intensity value in each small rectangular region can be obtained, and then the uniform distribution standard deviation of these fluorescence signal intensity values ​​can be calculated, and the uniform distribution standard deviation can be used as uniformity c). The uniformity of the first fluorescence after the chromatography reaction is completed is denoted as d (for example, the detection line region in the two-dimensional fluorescence signal image of the first fluorescence after the chromatography reaction is completed can be divided into 100 small rectangular regions of the same size, the fluorescence signal intensity value in each small rectangular region can be obtained, and then the uniform distribution standard deviation of these fluorescence signal intensity values ​​can be calculated, and the uniform distribution standard deviation can be used as uniformity d).

[0234] Then, the initial detection result X is corrected using the coating amount and uniformity of the captured material before and after the chromatography reaction, as well as the correction factor e related to the chromatography rate, to obtain the final detection result, denoted as Y5. For example, the final detection result Y5 = f(a, b, c, d, e, X) = X*a*a(bd) / [b*b*(ac)*e].

[0235] In the fourth embodiment, the immunochromatographic analysis method also obtains reference information of the immunochromatographic analysis reagent card. In this embodiment, the reference information is the reference chromatography speed V0. In some optional embodiments, the reference information may include at least the reference concentration information R0 of the first internal reference on the detection line and / or the reference fluorescence signal T0 of the first internal reference on the detection line and / or the reference chromatography speed V0 of the test strip and / or the item information and / or the calibration curve.

[0236] As an example, the initial detection result X can be corrected by combining the reference chromatography rate V0 and the chromatography rate-related correction factor e to obtain the final detection result, denoted as Y6. For example, Y6 = f(e, V0, X) = X*e / V0.

[0237] In the fifth embodiment, a computer-readable storage medium is provided, the computer-readable storage medium including a program that can be executed by a processor to implement the immunochromatographic analysis method in the first embodiment, the second embodiment, the third embodiment, the fourth embodiment and the extended embodiments thereof.

[0238] In a sixth embodiment, an immunochromatographic analyzer is provided, including an optical measurement device and a processor. The optical measurement device includes at least an optical imaging unit, which is used to acquire a two-dimensional fluorescence signal image of the immunochromatographic test strip. The processor is used to execute the computer-readable storage medium described in the fifth embodiment.

[0239] In some optional embodiments, the optical measurement device in the immunochromatographic analyzer further includes at least an optical scanning unit, which is used to acquire the one-dimensional fluorescence signal curve of the immunochromatographic test strip. In other optional embodiments, the optical imaging unit is selected from at least one of CCD and CMOS; the optical scanning unit is a photodetector.

[0240] In some other alternative implementations, the immunochromatographic analyzer also includes:

[0241] A sample loading module, for at least the automatic loading and unloading of sample holders or sample tubes; and / or a reagent card loading module, for at least the automatic loading of reagent cards; and / or an incubation module, for at least the carrying and incubation of reagent cards; and / or a temperature control unit, for at least the provision of the temperature conditions required for incubation; and / or a reagent card unloading module, for at least the automatic unloading of reagent cards.

[0242] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention without departing from the principles and spirit of the present invention.

Claims

1. An immunochromatographic analyzer characterized by comprising: The application relates to an immunochromatographic analyzer, comprising: a sample collection device, which is used for collecting a sample for preparing a test sample; a reagent supply device, which provides a reagent capable of reacting with the sample and participating in the preparation process of the test sample; a test sample transfer device, which is configured to transfer the test sample to a test strip of an immunochromatographic analysis reagent card; a light source assembly, which is configured to irradiate the test strip and is used for exciting a first fluorescent substance to generate first fluorescence and exciting a second fluorescent substance to generate second fluorescence; the first fluorescent substance comprises a first internal reference, which exists at least on a detection line of the test strip; an optical measurement device, which comprises an optical imaging unit and an optical scanning unit; the optical imaging unit is used for acquiring a two-dimensional fluorescence signal image of the immunochromatographic test strip; and the optical scanning unit is used for acquiring a one-dimensional fluorescence signal curve of the immunochromatographic test strip; a data processing unit, which is configured to correct a detection result by using at least part of information in the two-dimensional fluorescence signal image and / or at least part of information in the one-dimensional fluorescence signal curve, so as to obtain a corrected detection result.

2. An immunochromatographic assay device as claimed in claim 1, characterized in that Further comprising: a control unit, which is used for at least controlling the optical measurement device to acquire a first one-dimensional fluorescence signal curve C1 of the first fluorescence before the chromatographic reaction proceeds to the detection line; and / or, is used for at least controlling the optical measurement device to acquire a first two-dimensional fluorescence signal image I1 of the first fluorescence before the chromatographic reaction proceeds to the detection line; and / or, used for at least controlling the optical measurement device to acquire a second one-dimensional fluorescence signal curve C2 of the first fluorescence after the chromatographic reaction is completed; and / or, is used for at least controlling the optical measurement device to acquire a second two-dimensional fluorescence signal image I2 of the first fluorescence after the chromatographic reaction is completed.

3. The immunochromatographic analyzer according to claim 1 or 2, wherein: the second fluorescent substance comprises a second internal reference, which exists in the test sample and / or in a non-detection line area of the test strip before the chromatographic reaction; the immunochromatographic analyzer further comprises a control unit; the control unit is used for at least controlling the optical measurement device to acquire a third two-dimensional fluorescence signal image I3 of the second fluorescence during the chromatographic reaction; or, the control unit is used for at least controlling the optical measurement device to acquire a fourth two-dimensional fluorescence signal image I4 and a fifth two-dimensional fluorescence signal image I5 of the second fluorescence at two different moments during the chromatographic reaction.

4. The immunochromatographic analyzer according to any one of claims 1-3, wherein: the optical measurement device further comprises a first filter and a second filter; the first filter is used for filtering the first fluorescence; the second filter is used for filtering the second fluorescence; and / or, the optical imaging unit comprises at least one of a CCD and a CMOS; and the optical scanning unit comprises a photodetector.

5. An immunochromatographic assay as claimed in any one of claims 1 to 4, characterised in that, Further comprising: The information acquisition unit is configured to acquire reference information of the immunochromatographic assay reagent card, and the reference information comprises at least reference concentration information R0 of the first fluorescent substance on the detection line, and / or reference fluorescence signal T0 of the first fluorescent substance on the detection line, and / or reference chromatographic speed V0 of the test strip, and / or item information, and / or a calibration curve. The data processing unit is configured to correct the detection result by combining at least part of the reference information, at least part of the two-dimensional fluorescence signal image, and / or at least part of the one-dimensional fluorescence signal curve, to obtain a corrected detection result.

6. The immunochromatographic analyzer according to any one of claims 1-5, wherein the control unit is further configured to control the optical measurement device to acquire a third one-dimensional fluorescence signal curve C3 of the second fluorescence of the detection line region after the chromatographic reaction is completed and / or a sixth two-dimensional fluorescence signal image I6, and the third one-dimensional fluorescence signal curve C3 and / or the sixth two-dimensional fluorescence signal image I6 are used to acquire the detection result.

7. The immunochromatographic analyzer according to claim 5, wherein the data processing unit is further configured to acquire at least one detection result correction parameter by combining at least part of the reference information, at least part of the two-dimensional fluorescence signal images, and / or at least part of the one-dimensional fluorescence signal curves, and the at least one detection result correction parameter is used to correct the detection result to obtain a corrected detection result; and the detection result correction parameters acquired by the data processing unit comprise any one of a first capture coating amount correction factor related to the coating amount of the capture on the detection line before the chromatographic reaction, a second capture coating amount correction factor related to the coating amount of the capture on the detection line after the chromatographic reaction is completed, a first capture coating uniformity correction factor related to the coating amount of the capture on the detection line before the chromatographic reaction is completed, a second capture coating uniformity correction factor related to the coating amount of the capture on the detection line after the chromatographic reaction, an actual chromatographic speed correction factor related to the actual chromatographic speed, a combination of any two, a combination of any three, a combination of any four, or a combination of five. Further comprising: a result output unit, the result output unit is configured to output the detection result; and / or, the result output unit is configured to output at least one corrected detection result; 8. An immunochromatographic assay as claimed in any one of claims 1 to 7, characterised in that, and / or, the result output unit is configured to simultaneously output the detection result, at least one corrected detection result, and a correction parameter corresponding to the corrected detection result; and / or, the result output unit is configured to output the detection result and at least two corrected detection results for selection.

9. The immunochromatographic analyzer according to claim 8, wherein the result output unit comprises a display interface, and the display interface is configured to display the detection result and / or at least one corrected detection result. ​ ​ ​ ​ And / or, the display interface is used at least for displaying the detection result and at least two corrected detection results for selection, and in response to a selection instruction, the selected detection result or the corrected detection result is output as the final detection result.

10. The immuno-chromatography analyzer according to any one of claims 1-9, wherein, The light source assembly is used at least for providing first excitation light of a first wavelength and second excitation light of a second wavelength, the first excitation light is used for exciting the first fluorescent substance to generate first fluorescence, and the second excitation light is used for exciting the second fluorescent substance to generate second fluorescence; or, the light source assembly is used at least for providing third excitation light of a third wavelength, the third excitation light is used for exciting the first fluorescent substance to generate first fluorescence, and the third excitation light is used for exciting the second fluorescent substance to generate second fluorescence.

11. The immuno-chromatography analyzer according to any one of claims 8 or 9, wherein, The data processing unit is further used at least for converting the two-dimensional fluorescent signal image into a three-dimensional fluorescent signal image; The result output unit is further used at least for outputting the one-dimensional fluorescent signal curve and / or the two-dimensional fluorescent signal image and / or the three-dimensional fluorescent signal image.

12. An immunochromatographic assay as claimed in any one of claims 1 to 11, characterised in that, Further comprising: A sample loading module is used at least for automatic loading and unloading of a sample holder or a sample tube; and / or, a reagent card loading module is used at least for automatic loading of a reagent card; and / or, an incubation module is used at least for carrying and incubating the reagent card; and / or, a temperature control unit is used at least for providing temperature conditions required for incubation; and / or, a reagent card unloading module is used at least for automatic unloading of the reagent card.

13. An immunochromatographic assay method characterized by, Comprising: Providing an immuno-chromatography analysis reagent card to an immuno-chromatography analysis instrument, the immuno-chromatography analysis reagent card comprising a test strip, the test strip comprising at least a sample pad, an NC membrane, and a water absorption pad, the sample pad being located upstream of the NC membrane, the NC membrane being provided with a detection line, the detection line being coated with at least a first internal reference and a capture agent, the capture agent and the first internal reference being independently coated on the detection line, or the first internal reference being directly and / or indirectly combined with the capture agent; The light source irradiates the reagent card and excites the first internal reference to generate first fluorescence; Providing a test sample to the test strip of the immuno-chromatography analysis reagent card to start a chromatographic reaction; the test sample containing a tracer; The light source irradiates the reagent card and excites the tracer to generate second fluorescence; Obtaining a signal of the first fluorescence; The signal of the first fluorescence comprises: a two-dimensional fluorescent signal image of the first fluorescence and / or a one-dimensional fluorescent signal curve of the first fluorescence before the chromatographic reaction is performed to the detection line after the immuno-chromatography analysis reagent card is provided to the immuno-chromatography analysis instrument; and a two-dimensional fluorescent signal image of the first fluorescence and / or a one-dimensional fluorescent signal curve of first fluorescence after the chromatographic reaction is completed; Obtaining a signal of the second fluorescence; the signal of the second fluorescence comprises: a two-dimensional fluorescent signal image of the second fluorescence and / or a one-dimensional fluorescent signal curve of the second fluorescence after the chromatographic reaction is completed; and outputting a detection result by combining at least part of the information of the first fluorescence signal and at least part of the information of the second fluorescence signal.

14. An immunochromatographic assay method characterized by, The method comprises the following steps: Providing an immunochromatographic assay reagent card to an immunochromatographic analyzer, the immunochromatographic assay reagent card comprising a test strip, the test strip comprising at least a sample pad, an NC membrane, and a water absorption pad, the sample pad being located upstream of the NC membrane, the NC membrane being provided with a detection line, the detection line being coated with at least a capture agent, the capture agent being used for capturing at least a tracer in a chromatographic process; Providing a sample to the reagent card, the sample containing the tracer, so that the tracer in the sample chromatographs along the sample pad towards the water absorption pad on the test strip; Illuminating the reagent card with a light source and exciting the tracer to generate a second fluorescence; Obtaining a signal of the second fluorescence during the chromatographic reaction, comprising a seventh two-dimensional fluorescence signal image I7 of the second fluorescence; Obtaining a signal of the second fluorescence after the chromatographic reaction is completed, comprising an eighth two-dimensional fluorescence signal image I8 of the second fluorescence and / or a fourth one-dimensional fluorescence signal curve C4 of the second fluorescence; and outputting a detection result by combining at least part of the information of the signal of the second fluorescence during the chromatographic reaction and at least part of the information of the signal of the second fluorescence after the chromatographic reaction is completed.

15. The immunochromatographic assay method of claim 14, wherein: the detection result is output by combining at least part of the information of I7 and at least part of the information of I8; and / or, the detection result is output by combining at least part of the information of I7 and at least part the information of C4; and / or, the detection result is output by combining at least part of the information of I 7, at least part of the information of I8, and at least part of the information of C4.

16. The immunochromatographic assay method of claim 14, wherein: the detection line is further coated with at least a first internal reference, the capture agent and the first internal reference being independently coated on the detection line, or the first internal reference being directly and / or indirectly combined with the capture agent; the reagent card is illuminated with a light source to excite the first internal reference to generate a first fluorescence; obtaining a signal of the first fluorescence before the chromatographic reaction reaches the detection line, comprising a ninth two-dimensional fluorescence signal image I9 of the first fluorescence and / or a fifth one-dimensional fluorescence signal curve C5 of the first fluorescence; and outputting a detection result by combining at least part of the information of the signal during the chromatographic reaction, at least part of the information of the signal after the chromatographic reaction is completed, and at least part of the information of the signal of the first fluorescence before the chromatographic reaction reaches the detection line; or obtaining a signal of the first fluorescence after the chromatographic reaction is completed, comprising a twelfth two-dimensional fluorescence signal image I10 of the first fluorescence and / or a sixth one-dimensional fluorescence signal curve C6 of the first fluorescence; and outputting a detection result by combining at least part of the information during the chromatographic reaction, at least part of the information after the chromatographic reaction is completed, and at least part of the information of the first fluorescence before the chromatographic reaction reaches the detection line. ​ Or, the signal of the first fluorescence before the detection line is obtained, including: the ninth two-dimensional fluorescence signal image I9 of the first fluorescence and / or the fifth one-dimensional fluorescence signal curve C5 of the first fluorescence; the signal of the first fluorescence after the completion of the chromatography reaction is obtained, including: the twelfth two-dimensional fluorescence signal image I10 of the first fluorescence and / or the sixth one-dimensional fluorescence signal curve C6 of the first fluorescence; at least the part of the information of the signal of the second fluorescence during the chromatography reaction, the part of the information of the signal of the second fluorescence after the completion of the chromatography reaction, the part of the information of the signal of the first fluorescence before the detection line and the part of the information of the signal of the first fluorescence before the detection line are combined to output the detection result.

17. The immuno-chromatographic analysis method according to any one of claims 14-16, wherein the signal of the second fluorescence during the chromatography reaction further comprises: The signal of the second fluorescence at the first time during the chromatography reaction is obtained, including: the eleventh two-dimensional fluorescence signal image I11 of the second fluorescence at the first time; the signal of the second fluorescence at the second time during the chromatography reaction is obtained, including: the tenth two-dimensional fluorescence signal image I12 of the second fluorescence at the second time; and at least the part of the information in the fifth two-dimensional fluorescence signal image I11 and the part of the information in the sixth two-dimensional fluorescence signal image I12 are used to output the detection result. A program is included, which can be executed by a processor to implement the immuno-chromatographic analysis method according to any one of claims 13-17.

18. A computer-readable storage medium, characterized in that, An optical measurement device is included, which at least comprises an optical imaging unit used to obtain the two-dimensional fluorescence signal image of the immuno-chromatographic test strip, and a processor used to execute the computer readable storage medium according to claim 18.

19. An immunochromatographic analyzer characterized by comprising:

20. The immuno-chromatographic analyzer according to claim 19, wherein the optical measurement device further comprises an optical scanning unit used to obtain the one-dimensional fluorescence signal curve of the immuno-chromatographic test strip; the optical imaging unit is selected from at least one of a CCD and a CMOS; and the optical scanning unit is a photoelectric detector. ​ ​

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