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Fluorescence immunochromatography test strip for quantitatively detecting anti-PLA2R antibody and preparation method of fluorescence immunochromatography test strip

A fluorescence immunochromatography and PLA2R technology, which is applied in the field of fluorescence immunochromatography test strips and their preparation, can solve the problem of inability to quickly and accurately detect the content of anti-PLA2R antibodies, high technical requirements for testing personnel, and inability to guarantee 100% response. and other problems, to achieve the effect of short detection time, low personnel quality requirements and simple structure

Pending Publication Date: 2022-05-06
迪亚莱博(张家港)生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the clinically recognized PLA2R detection method mainly uses the indirect immunofluorescence kit of Oumeng Company to test the anti-PLA2R antibody content in serum, but the operation method of the kit is relatively complicated, the detection cycle is long, and it is difficult for the technical personnel of the detection personnel. High requirements, this method cannot meet the requirements of rapid and accurate detection of anti-PLA2R antibody content, and has many limitations
[0005] The existing patent CN104849452A sprays quantum dots coupled with anti-human IgG antibody on the binding pad, and the nitrocellulose membrane is coated with PLA2R antigen as the detection line, which has the problems of insufficient sensitivity and inaccurate detection results
The existing patent CN111198273A is also in the form of anti-human IgG labeling and PLA2R antigen coating, which also has the problem of insufficient sensitivity and inaccurate detection results
The existing patent CN111413506A has two binding pads, the structure is relatively complex, and mass production is difficult, and it needs to biotinylate PLA2R, which is complicated to operate and consumes raw materials. 100% reaction of fluorescent microspheres, easy to cause signal loss

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  • Fluorescence immunochromatography test strip for quantitatively detecting anti-PLA2R antibody and preparation method of fluorescence immunochromatography test strip
  • Fluorescence immunochromatography test strip for quantitatively detecting anti-PLA2R antibody and preparation method of fluorescence immunochromatography test strip
  • Fluorescence immunochromatography test strip for quantitatively detecting anti-PLA2R antibody and preparation method of fluorescence immunochromatography test strip

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Embodiment 1

[0052] This embodiment provides a fluorescent immunochromatographic test strip for the quantitative detection of anti-PLA2R antibodies, the structure of which is as follows: figure 1 As shown, it includes sample pad 1, bottom plate 2, binding pad 3, detection line 4, nitrocellulose membrane 5, quality control line 6, and absorbent paper 7. Mouse anti-human erythrocyte antibody is sealed on sample pad 1, and binding pad 3 is sealed. The PLA2R protein labeled with fluorescent microspheres and the rabbit IgG antibody labeled with fluorescent microspheres were sprayed; the antibody coated on the detection line 4 was mouse anti-human IgG antibody; the antibody coated on the quality control line 6 was goat anti-rabbit IgG antibody. Among them, the material of the base plate 2 is PVC, the material of the sample pad 1 and the binding pad 3 are glass cellulose membranes, the fluorescent microspheres are europium chelated fluorescent microspheres, purchased from Thermo, and the modified ...

Embodiment 2

[0102] Get 70 clinical serum samples, use the fluorescent immunochromatography test strip of embodiment 1 to quantitatively detect anti-PLA2R antibody to detect, and the detection method is:

[0103](1) Use the fluorescent immunoassay analyzer to read the ID card of the pre-made PLA2R, and save the standard curve.

[0104] (2) Take 5 μL of the positive sample, mix it with 95 μL of the diluent, take 80 μL of the mixed solution and add it to the sample hole of the PLA2R test strip. The diluent is 50 mM Tris buffer with a pH value of 8.5.

[0105] (3) After waiting for 8 minutes, insert the fluorescent immunoassay analyzer to test the T value and C value, and calculate the anti-PLA2R antibody concentration by substituting the T / C value into the standard curve.

[0106] The results were compared with the test results of the industry-recognized indirect immunofluorescence kit of Oumeng Company, and the results are shown in Table 5.

[0107] table 5

[0108] quantity ...

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Abstract

The invention provides a fluorescence immunochromatography test strip for quantitatively detecting an anti-PLA2R antibody and a preparation method thereof, the fluorescence immunochromatography test strip comprises a bottom plate, and a sample pad, a combination pad, a nitrocellulose membrane and absorbent paper which are sequentially carried on the bottom plate, the conjugate pad is coated with a PLA2R fluorescent microsphere conjugate and a rabbit IgG fluorescent microsphere conjugate, the nitrocellulose membrane is coated with a mouse anti-human IgG antibody as a detection line, and the nitrocellulose membrane is also coated with goat anti-rabbit IgG as a quality control line. The fluorescence immunochromatography test strip disclosed by the invention is high in sensitivity and accuracy, extremely small in dosage of required clinical samples and simple to operate, and signal detection and concentration value conversion can be realized by only one fluorescence immunoassay analyzer.

Description

technical field [0001] The invention relates to the technical field of immunofluorescence detection, in particular to a fluorescent immunochromatographic test strip for quantitatively detecting anti-PLA2R antibodies and a preparation method thereof. Background technique [0002] Phospholipase A2 receptor (PLA2R) is a transmembrane glycoprotein belonging to the mannitol receptor family. Its molecular mass is 185KD. It was first discovered in human normal podocytes, and its main expression is in glomerular podocytes. . Serum anti-PLA2R antibody in patients with idiopathic membranous nephropathy binds to PLA2R antigen, and the two form an in situ immune complex that deposits on the glomerular basement membrane, causing podocyte damage, glomerular basement membrane thickening, and glomerular basement membrane damage. Therefore, the clinical detection of PLA2R in renal tissue is of great significance for the diagnosis, differential diagnosis, disease assessment, curative effect ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/58G01N33/558G01N33/52
CPCG01N33/6893G01N33/6872G01N33/558G01N33/582G01N33/52G01N2333/705G01N2800/347Y02A50/30
Inventor 徐国新姚辉
Owner 迪亚莱博(张家港)生物科技有限公司
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