A reconstituted solution of fluorescent microsphere-labeled antibody and its use in the preparation of a fluorescent microsphere-labeled immunochromatographic test strip for lipoarabinomannan

Abstract

The application discloses a reconstituted solution of fluorescent microsphere labeled antibody and application of the reconstituted solution in preparation of a liparabinomannan fluorescent microsphere immunochromatographic test strip. The reconstituted solution of the fluorescent microsphere labeled antibody comprises HEPES buffer, bovine serum albumin, trehalose, sucrose and Pluronic L64. The reconstituted solution of the fluorescent microsphere labeled antibody can significantly enhance the stability of the fluorescent microsphere labeled antibody, effectively avoid the decrease of activity and detection sensitivity of the fluorescent microsphere labeled antibody in a short-term storage process, and reduce batch difference, and has a wide application prospect in large-scale production of the fluorescent microsphere immunochromatographic test strip.

Description

Technical Field

[0001] This invention relates to the technical field of immunoassay, specifically to a reconstituted solution of fluorescent microsphere-labeled antibody and its application in the preparation of lipoarabinomannan fluorescent microsphere immunochromatographic test strips. Background Technology

[0002] Fluorescent immunochromatography is a novel membrane detection technology based on antigen-antibody specific immune reactions. This technology uses a strip-shaped chromatographic material immobilized with a detection line (coated antibody or antigen) and a control line as the stationary phase, and the test solution as the mobile phase. Fluorescently labeled antibodies or antigens are immobilized on the binding pads, and the analyte moves along the chromatographic strip via capillary action. For large molecular antigens (proteins, viruses, pathogens, etc.) with multiple antigenic determinants, a "sandwich" type double-antibody immunochromatographic method is typically used: "antibody-antigen-fluorescent antibody." In this method, the analyte first binds to the fluorescently labeled antibody under the action of the mobile phase, and then binds to the coated antibody when it reaches the detection line, forming a double-antibody sandwich.

[0003] Quantum dot fluorescent microspheres are micron / nanoscale fluorescent particles formed by embedding semiconductor nanocrystals into polymers or inorganic matrices through assembly processes. The particle size is typically 100–400 nm. The core of these microspheres is encapsulated with inorganic materials or polymers to enhance stability and reduce environmental interference. The surface of these quantum dot fluorescent microspheres is modified with hydroxyl, amino, and other groups, facilitating coupling with biomolecules such as antibodies and nucleic acids for targeted detection.

[0004] Fluorescent microsphere-labeled antibodies typically require preservation in reconstitution solutions. However, fluorescent microspheres are highly sensitive to pH and ionic strength, making the composition of the reconstitution solution crucial. An unsuitable reconstitution solution can lead to microsphere aggregation or fluorescence quenching. Furthermore, fluorescent microsphere-labeled antibodies sometimes require storage for a certain period during production before use, which can further increase the likelihood of aggregation and quenching of the fluorescent groups during storage, resulting in decreased stability and significantly reduced detection sensitivity. Currently used reconstitution solutions offer insufficient protection for fluorescent microsphere-labeled antibodies, easily causing fluorescence decay or decline within a certain timeframe, indicating poor stability and hindering large-scale production and preservation. Therefore, developing a suitable reconstitution solution for fluorescent microsphere-labeled antibodies is of great significance for fluorescence immunochromatographic detection. Summary of the Invention

[0005] To overcome the aforementioned defects and shortcomings in the prior art, this invention provides a reconstituted solution of fluorescent microsphere-labeled antibody and its application in the preparation of lipoarabinomannan fluorescent microsphere immunochromatographic test strips.

[0006] The first object of the present invention is to provide the use of a composition in the preparation of a reconstitution solution of a fluorescent microsphere-labeled antibody.

[0007] A second objective of this invention is to provide a reconstitution solution of a fluorescent microsphere-labeled antibody.

[0008] The third objective of this invention is to provide a fluorescent microsphere immunochromatographic test strip.

[0009] A fourth objective of this invention is to provide the application of the above-described fluorescent microsphere immunochromatographic test strip in the preparation of products for detecting Mycobacterium tuberculosis and / or diagnosing tuberculosis.

[0010] This invention claims protection for the following: The use of a composition in the preparation of a reconstitution solution of a fluorescent microsphere-labeled antibody, said composition comprising bovine serum albumin, trehalose, sucrose, Pluronic L64, and a buffer solution; The bovine serum albumin has a mass concentration of 0.8–1.2%, the trehalose has a mass concentration of 1.8–2.2%, the sucrose has a mass concentration of 2.8–3.2%, and the Pluronic L64 has a mass concentration of 0.5–1%.

[0011] Preferably, the mass concentration of bovine serum albumin is 1%, the mass concentration of trehalose is 2%, the mass concentration of sucrose is 3%, and the mass concentration of Pluronic L64 is 0.5-1%.

[0012] Preferably, the buffer solution is a HEPES buffer solution.

[0013] Preferably, the concentration of the HEPES buffer is 8–12 mM.

[0014] More preferably, the concentration of the HEPES buffer is 10 mM.

[0015] Preferably, the antibody is a lipoarabinomannan antibody.

[0016] A reconstitution solution of a fluorescent microsphere-labeled antibody, the reconstitution solution containing bovine serum albumin, trehalose, sucrose, Pluronic L64 and buffer solution; The bovine serum albumin has a mass concentration of 0.8–1.2%, the trehalose has a mass concentration of 1.8–2.2%, the sucrose has a mass concentration of 2.8–3.2%, and the Pluronic L64 has a mass concentration of 0.5–1%.

[0017] Preferably, the mass concentration of bovine serum albumin is 1%, the mass concentration of trehalose is 2%, the mass concentration of sucrose is 3%, and the mass concentration of Pluronic L64 is 0.5-1%.

[0018] Preferably, the buffer solution is a HEPES buffer solution.

[0019] Preferably, the concentration of the HEPES buffer is 8–12 mM.

[0020] More preferably, the concentration of the HEPES buffer is 10 mM.

[0021] Preferably, the pH of the reconstituted solution is 7.4 to 7.6.

[0022] More preferably, the pH of the reconstituted solution is 7.5.

[0023] A fluorescent microsphere immunochromatographic test strip, wherein the fluorescent microsphere immunochromatographic test strip comprises a fluorescent microsphere pad, and the preparation method of the fluorescent microsphere pad includes the following steps: S1. Label the lipoarabinomannan antibody with fluorescent microspheres to obtain the labeled antibody; S2. Dilute the labeled antibody with any of the above-mentioned reconstitution solutions to obtain a labeled antibody diluent. Spray the labeled antibody diluent evenly onto a glass cellulose membrane and dry it to obtain a fluorescent microsphere pad.

[0024] The above-mentioned fluorescent microsphere immunochromatographic test strips are used in the preparation of products for detecting Mycobacterium tuberculosis and / or diagnosing tuberculosis.

[0025] Compared with the prior art, the present invention has the following beneficial effects: This invention discloses a reconstitution solution for fluorescent microsphere-labeled antibodies and its application in the preparation of lipoarabinomannan fluorescent microsphere immunochromatographic test strips. In the reconstitution solution, Pluronic L64 exhibits a good synergistic effect with carbohydrates and bovine serum albumin, which can slow down the fluorescence quenching and aggregation of fluorescent microspheres, making the microspheres more stable and beneficial for long-term storage and use. Therefore, the reconstitution solution of this invention can effectively reduce the fluorescence quenching rate of fluorescent microspheres and significantly improve the stability of fluorescent microsphere-labeled antibodies, allowing them to be stably stored at 2–8°C for at least 20 days, effectively avoiding the decrease in activity and detection sensitivity of fluorescent microsphere-labeled antibodies during short-term storage. Applying the reconstitution solution of this invention to the preparation of fluorescent microsphere immunochromatographic test strips is not only simple to operate but also makes the fluorescent microsphere-labeled antibodies more stable, suitable for storage after industrial production, and can effectively control batch-to-batch variations. It has broad application prospects in the large-scale production of lipoarabinomannan fluorescent microsphere immunochromatographic test strips. Detailed Implementation

[0026] The present invention will be further illustrated below with reference to specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods, and equipment used in the present invention are conventional reagents, methods, and equipment in this technical field.

[0027] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0028] Fluorescent microspheres: purchased from Bangs Laboratories, with a particle size of 0.2 μm; Microsphere dilution buffer: 0.02 M phosphate buffer containing 0.05% Tween-20 at pH 8.0; Labeling buffer: 0.02 M borate buffer, pH 7.5; Blocking agent: 0.05% borate buffer (w / v) containing 5% sodium casein (w / v), pH 7.5; Coating buffer: 20 mM PBS buffer, pH 7.4; Sample pad treatment solution / conjugation pad treatment solution: 0.05 M Tris buffer containing 2.0% trehalose, 5% sucrose and 0.2% Tetronic 1307 (S9) at pH 7.5.

[0029] Example 1: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This embodiment provides a reconstitution solution of fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 0.5% Pluronic L64 (S23) at a mass concentration of 7.5.

[0030] The fluorescent microsphere-labeled antibody is reconstituted using a reconstitution solution of the fluorescent microsphere-labeled antibody, and then a fluorescent microsphere immunochromatographic test strip is prepared. The preparation method of the fluorescent microsphere immunochromatographic test strip includes the following steps: 1. Preparation of activator: Prepare 0.025 mol / L MES buffer solution with ultrapure water, and dissolve NHS and EDC separately with MES buffer solution to obtain NHS solution and EDC solution with a concentration of 20 mg / mL. Prepare fresh solution and use immediately. 2. Take 100 μL of microsphere dilution solution from each of two tubes, and add fluorescent microspheres to each tube to make the final concentration of fluorescent microspheres 10 mg / mL, thus obtaining a fluorescent microsphere solution; 3. Add 5 μL of EDC solution to 100 μL of fluorescent microsphere solution and mix well. Then add 5 μL of NHS solution and mix thoroughly. Place in a mixer at 25℃ for 30 min to activate. After activation, centrifuge at 11600 g for 15 min, discard the supernatant, and resuspend the precipitate in 100 μL of labeling buffer and mix well to obtain the resuspension of fluorescent microspheres. 4. Add lipoarabinomannan (LAM) monoclonal antibody (LAM-coated antibody, brand: creative Diagnostics, catalog number: CABT-L0374Y) to one tube of fluorescent microsphere resuspension, and add DNP-BSA antibody (brand: Nanjing Fuxiao Biotechnology, catalog number: DNP-T112501M) to the other tube of fluorescent microsphere resuspension, so that the final antibody concentration is 200 μg / mL, and perform the coupling reaction for 2 h; 5. After the coupling reaction, add 100 μL of blocking agent to block the excess sites of the fluorescent microspheres, then centrifuge at 11600 g for 15 min, discard the supernatant, add the reconstitution solution of the fluorescent microsphere labeled antibody to the precipitate, mix well to obtain the fluorescent microsphere labeled antibody dilution solution (fluorescent microsphere dilution solution of LAM monoclonal labeled antibody and fluorescent microsphere dilution solution of DNP-BSA antibody), place at 2-8℃ for 0 days, 5 days, 10 days, 15 days and 20 days respectively, then dilute with the reconstitution solution at a volume ratio of 1:1, spray on a 7 mm wide glass cellulose membrane, place in an oven at 37℃ for 1 h to obtain fluorescent microsphere pads of different groups; 6. Preparation of the detection line (T line) and control line (C line): The T line antibody (LAM detection antibody, brand: creativeDiagnostics, catalog number: CABT-L0375Y) and the C line antibody (DNP-BSA antibody, brand: Nanjing Fuxiao Biotechnology, catalog number: DNP-T112505R) were diluted with coating buffer and coated onto the NC membrane. The coating concentration of the T line antibody was 1.2 mg / mL and the coating concentration of the C line antibody was 1.0 mg / mL. Using a membrane scribing instrument, a straight line was uniformly scribed in the middle of the NC membrane at a coating amount of 1.1 μL / cm to form the detection line (T line) and the control line (C line). Then, the membrane was placed in an oven and dried at 37℃ for 1 h. 7. Sample pad preparation: Cut glass cellulose membrane into strips 1.7 cm wide, immerse the cut glass cellulose membrane in sample pad treatment solution, and then dry it in an oven at 37°C for 3 h to obtain the sample pad; 8. Absorbent pad: Cut absorbent paper into strips 1.7 cm wide and dry at 37℃ for 3 hours to obtain an absorbent pad; 9. Assemble the sample pad, fluorescent microsphere pad, NC membrane, and absorbent pad onto the PVC base plate in sequence, cut them into 3 mm wide test strips, and assemble them into a test card to obtain the fluorescent microsphere immunochromatographic test strip.

[0031] Example 2: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This embodiment provides a reconstitution solution of fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1.0% Pluronic L64 (S23) at a mass concentration of 7.5.

[0032] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0033] Comparative Example 1: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 0.5% BSA, 1% trehalose and 3% sucrose, with a pH of 7.5.

[0034] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0035] Comparative Example 2: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose and 3% sucrose, with a pH of 7.5.

[0036] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0037] Comparative Example 3: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1% Tween-20 (S19) at a mass concentration of 7.5.

[0038] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0039] Comparative Example 4: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1% Tetronic 1307 (S9) at a mass concentration of 7.5.

[0040] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0041] Comparative Example 5: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1% Triton X-100 (S14) at a mass concentration of 7.5.

[0042] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0043] Comparative Example 6: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1% Tween-80 (S20) at a mass concentration of 7.5.

[0044] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0045] Comparative Example 7: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 1% BRIJ 35 (S21) at a mass concentration of 7.5.

[0046] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0047] Comparative Example 8: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.5% Tris buffer, 0.9% sodium chloride, 1% bovine serum albumin, 10% sucrose, 5% trehalose, and 0.1% Tetronic 1307 (S9), with a pH of 7.5.

[0048] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0049] Comparative Example 9: A reconstituted solution of a fluorescent microsphere-labeled antibody and a fluorescent microsphere immunochromatographic test strip for detecting lipoarabinomannan. This comparative example provides a reconstitution solution of a fluorescent microsphere-labeled antibody, comprising the following components: 0.01 M HEPES buffer, 1% BSA, 2% trehalose, 3% sucrose and 2.0% Pluronic L64 (S23) at a mass concentration of 7.5.

[0050] The preparation method of the fluorescent microsphere immunochromatographic test strip was carried out in accordance with Example 1.

[0051] Test Example 1: Sensitivity Detection I. Experimental Methods Fluorescent microsphere-labeled antibodies (stored for 0 days) from the reconstituted solutions of Examples 1-2 and Comparative Examples 1-9 were used to prepare fluorescent microsphere immunochromatographic test strips. The fluorescent microsphere immunochromatographic test strips were then used to detect 0.5 ng / mL LAM antigen samples. Each sample was tested 10 times. The T-line signal value was quantitatively detected using a fluorescence immunoassay analyzer to determine the sample concentration signal. The mean, standard deviation (SD), and coefficient of variation (CV) were calculated. A CV of less than 10% is acceptable for use.

[0052] II. Experimental Results The results are shown in Table 1. The fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1 and 2 were used to prepare fluorescent microsphere immunochromatographic test strips after being stored for 0 days. All of them were able to effectively detect LAM antigen samples (CV value less than 10%), with a sensitivity of 0.5 ng / mL.

[0053] The CV values ​​of the fluorescent microsphere-labeled antibodies prepared from the reconstituted solutions of Comparative Examples 1–9 when detecting LAM antigen samples were all greater than 10% after 0 days of storage, indicating that the test strips of Comparative Examples 1–9 could not effectively detect 0.5 ng / mL LAM antigen samples.

[0054] Table 1. Fluorescence values ​​of different groups of fluorescent microsphere immunochromatographic test strips.

[0055] Test Example 2: Stability Test I. Experimental Methods The fluorescent microsphere immunochromatographic test strips of Examples 1-2 and Comparative Examples 1-9 were used to detect 0.5 ng / mL and 10 ng / mL LAM antigen samples. Each sample was tested 10 times. The T-line signal value was quantitatively detected using a fluorescence immunoassay analyzer to determine the sample concentration signal. The mean, standard deviation (SD), and coefficient of variation (CV) were calculated. A CV of less than 10% is acceptable for use.

[0056] II. Experimental Results Table 2 shows the fluorescence values ​​of different groups of fluorescent microsphere immunochromatographic test strips when detecting 0.5 ng / mL LAM samples. The relative deviation of the fluorescent microsphere-labeled antibody prepared after 20 days of storage in the reconstituted solutions of Examples 1-2 for detecting 0.5 ng / mL LAM antigen samples was only about 5% compared with that at 0 days. This indicates that the reconstituted solutions of Examples 1-2 can reduce environmental interference during the storage of fluorescent microsphere-labeled antibodies, enhance the stability of fluorescent microsphere-labeled antibodies, and will not have a significant impact on the sensitivity of the test strips.

[0057] The relative deviations of the fluorescent microsphere-labeled antibodies prepared from the reconstituted solutions of Comparative Examples 1–9 after 20 days of storage for detecting 0.5 ng / mL LAM antigen samples compared to those after 0 days reached 27.23%–39.88%. This indicates that with prolonged storage time, the fluorescent microsphere-labeled antibodies of Comparative Examples 1–9 suffer from insufficient protection due to the reconstituted solutions, leading to a decrease in the stability and detection sensitivity of the fluorescent microspheres, making effective detection impossible.

[0058] Table 2. Fluorescence values ​​of different groups of fluorescent microsphere immunochromatographic test strips when detecting 0.5 ng / mL LAM samples.

[0059]

[0060] The fluorescence values ​​of different groups of fluorescent microsphere immunochromatographic test strips when detecting 10 ng / mL LAM samples are shown in Table 3. The fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1, 2, 3, and 9, when prepared into fluorescent microsphere immunochromatographic test strips after being stored for 0 days, can all effectively detect 10 ng / mL LAM antigen samples.

[0061] However, the detection CV values ​​of the fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Comparative Examples 3 and 9, respectively, were greater than 10% after 5 days and 15 days of storage, indicating that the test strips in these two groups could not effectively detect LAM antigen samples after the fluorescent microsphere-labeled antibodies had been stored for a period of time. In contrast, the detection CV value of the fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1 and 2, after 20 days of storage, was only about 4.8%, with a relative deviation of 8.13% to 9.62%, indicating that the fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1 and 2 have strong stability.

[0062] Table 3. Fluorescence values ​​of different groups of fluorescent microsphere immunochromatographic test strips when detecting 10 ng / mL LAM samples.

[0063]

[0064] Test Example 3: Production Batch Difference I. Experimental Methods Fluorescent microsphere-labeled antibodies (stored in reconstituted solutions of Examples 1-2 and Comparative Examples 1-9, stored for 0 days) were used to prepare fluorescent microsphere immunochromatographic test strips. Three different batches of test strips were prepared at 3-day intervals. These test strips were then used to test 10 ng / mL LAM antigen samples, and the T-line signal value was quantitatively detected using a fluorescence immunoassay analyzer. Each batch was performed in triplicate, and the mean value of each batch was calculated to determine the inter-batch relative deviation (R). The calculation method was as follows:

[0065] II. Experimental Results The results are shown in Table 4. The batch-to-batch relative deviation of fluorescent microsphere-labeled antibodies (stored for 0 days) prepared using the reconstituted solutions of Examples 1-2 was only 8.3% to 8.9%, which was lower than that of Comparative Examples 1-9. This indicates that the fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1-2 have better stability and are more suitable for commercial large-scale production of fluorescent microsphere immunochromatographic test strips.

[0066] Table 4. Results of inter-batch differences in production

[0067] Test Example 4: Clinical Sample Testing I. Experimental Methods Fluorescent microsphere-labeled antibodies (stored for 0 days) from the reconstituted solutions of Examples 1-2 and Comparative Examples 1-9 were used to prepare fluorescent microsphere immunochromatographic test strips. Clinical Mycobacterium tuberculosis liquid culture medium was then diluted to 10 ng / mL with culture medium. 80-100 μL of the Mycobacterium tuberculosis liquid culture medium to be tested was added to the sample well of the test strip using a dropper or pipette. Blank culture medium was used as a control. Two replicates were performed for each group, and the strips were incubated at room temperature for 15 min to determine the results. The T-line signal value was quantitatively detected using a fluorescence immunoassay analyzer, and the mean value was calculated.

[0068] II. Experimental Results The results are shown in Table 5. The fluorescent microsphere-labeled antibodies preserved in the reconstituted solutions of Examples 1 and 2 showed better stability and better detection sensitivity for clinical samples.

[0069] Table 5 Clinical Sample Test Results

[0070] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The use of a composition in the preparation of a reconstitution solution of fluorescent microsphere-labeled antibodies, characterized in that, The composition comprises bovine serum albumin, trehalose, sucrose, Pluronic L64, and a buffer solution; The bovine serum albumin has a mass concentration of 0.8–1.2%, the trehalose has a mass concentration of 1.8–2.2%, the sucrose has a mass concentration of 2.8–3.2%, and the Pluronic L64 has a mass concentration of 0.5–1%.

2. The application according to claim 1, characterized in that, The buffer solution is HEPES buffer.

3. The application according to claim 2, characterized in that, The concentration of the HEPES buffer solution is 8–12 mM.

4. The application according to claim 1, characterized in that, The antibody is a lipoarabinomannan antibody.

5. A reconstituted solution of a fluorescent microsphere-labeled antibody, characterized in that, The reconstitution solution contains bovine serum albumin, trehalose, sucrose, Pluronic L64, and buffer solution; The bovine serum albumin has a mass concentration of 0.8–1.2%, the trehalose has a mass concentration of 1.8–2.2%, the sucrose has a mass concentration of 2.8–3.2%, and the Pluronic L64 has a mass concentration of 0.5–1%.

6. The complex solution according to claim 5, characterized in that, The buffer solution is HEPES buffer.

7. The complex solution according to claim 6, characterized in that, The concentration of the HEPES buffer solution is 8–12 mM.

8. The complex solution according to claim 5, characterized in that, The pH of the reconstituted solution is 7.4–7.

6.

9. A fluorescent microsphere immunochromatographic test strip, characterized in that, The fluorescent microsphere immunochromatographic test strip comprises a fluorescent microsphere pad, and the preparation method of the fluorescent microsphere pad includes the following steps: S1. Label the lipoarabinomannan antibody with fluorescent microspheres to obtain the labeled antibody; S2. Dilute the labeled antibody with the reconstitution solution according to any one of claims 5 to 8 to obtain a labeled antibody diluent, uniformly spray the labeled antibody diluent onto a glass cellulose membrane, and obtain fluorescent microspheres after drying.

10. The use of the fluorescent microsphere immunochromatographic test strip of claim 9 in the preparation of products for detecting Mycobacterium tuberculosis and / or diagnosing tuberculosis.

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