Immunochromatography reagent strip for AMH fluorescent quantitative detection and preparation method of immunochromatography reagent strip

A technology of fluorescent quantitative detection and immunochromatographic test paper, which is applied in the field of immunodiagnosis, can solve the problems of many influencing factors, high price, false positives, etc., and achieve the effect of ensuring accuracy, high sensitivity and simple operation

Inactive Publication Date: 2017-09-22
SHENZHEN YHLO BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA method has many detection steps, takes a long time, and there are many influencing factors in the operation process, which can easily cause false positive and false negative results
Therefore, it is gradually replaced by chemiluminescence method, but this kind of method is a fully closed system, which is expensive and requires special training for instrument users. The maintenance and testing costs are high, and it is not suitable for single-person and small-batch testing. It is not conducive to AMH at present. Extensive testing in China
[0005] In view of the fact that there is no fast and accurate method for quantitatively detecting AMH at present, the purpose of the present invention is to provide a kind of immunochromatographic test strip that can be used for rapidly quantitatively detecting AMH, for evaluating and monitoring women's ovarian reserve function, said reagent It has the advantages of simple operation, convenience, economy, accurate quantification, etc., and is more suitable for extensive use in various medical institutions.

Method used

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  • Immunochromatography reagent strip for AMH fluorescent quantitative detection and preparation method of immunochromatography reagent strip

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Preparation of AMH fluorescent immunochromatographic test strips:

[0032] (1) Preparation of fluorescent microsphere-labeled protein

[0033]Take 0.1 mL of 10% fluorescent microspheres, centrifuge at 15,000 rpm for 15 minutes for the first time, adjust the concentration of the precipitate to 1% with 50 mM pH 6.5 phosphate buffer, and ultrasonically disperse; add carbon dioxide at a final concentration of 2 mg / mL imine (EDC), mix well, then add N-hydroxysuccinimide (NHS) with a final concentration of 5 mg / mL, mix well; incubate at room temperature for 20 minutes and then centrifuge at 15,000 rpm for a second time for 15 minutes. Dissolve in 50 mM pH 6.5 phosphate buffer. Ultrasonic disperse the reconstituted fluorescent microspheres, add 0.2 mg AMH monoclonal antibody and 0.1 mg avidin into two tubes respectively, mix well, rotate and mix at room temperature for 2 hours, centrifuge for the third time at 15,000 rpm for 15 minutes, and precipitate The material was redis...

Embodiment 2

[0042] Quantitative detection of anti-Müllerian hormone (AMH) concentration in blood samples by fluorescence immunochromatography

[0043] (1) Standard curve drawing

[0044] The AMH antigen was prepared with negative plasma to 25 ng / mL, 10 ng / mL, 5 ng / mL, 1.0 ng / mL, 0.5 ng / mL, 0.1 ng / mL, 0 ng / mL, using the same batch of reagents, Each concentration point was tested 6 times. Take the fluorescence intensity ratio of the detection line (T band) and the quality control line (C band) as the ordinate, and the AMH reference substance concentration as the abscissa, establish an equation and fit it into a standard curve, and write the standard curve information into In the ID chip.

[0045] (2) Detection of samples:

[0046] Take out the test strip from the kit, tear open the aluminum foil bag, lay the test strip flat, and equilibrate for 5 minutes. Take 100 μL of sample and add it to the sample well, and react at room temperature for 15 minutes in the dark. Insert the ID chip int...

Embodiment 3

[0050] Please refer to figure 1 , an immunochromatographic test strip for fluorescent quantitative detection of AMH, the immunochromatographic test strip includes a PVC base plate 7, and the PCV base plate 7 is sequentially provided with a sample pad 1, a marker pad 2, and a coating film 3. Absorbent paper 4, the marking pad 2 is connected to the sample pad 1, the coating film 3 is connected to the marking pad 2, the absorbent paper 4 is connected to the coating film 3 are connected; the AMH monoclonal antibody labeled with fluorescent microspheres and avidin labeled with fluorescent microspheres are sprayed on the marker pad 2; a detection line 5 and a quality control line 6 are provided on the coating film 3, and the detection line 5 and the quality control line 6 are separated by 4-8 mm, and the detection line 5 is coated with another AMH monoclonal antibody at a different epitope from the AMH monoclonal antibody labeled with the fluorescent microspheres, and the quality co...

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Abstract

An immunochromatographic test strip for fluorescent quantitative detection of AMH and a preparation method thereof. The invention belongs to the technical field of immunodiagnosis. The purpose of the invention is to address the deficiencies in the prior art. The invention adopts the following technical solutions: Immunochromatographic test strips for quantitative detection of AMH are composed of sample pads, marking pads, coating films, and absorbent papers that are sequentially lapped on a PVC bottom plate. The marking pads are sprayed with avidin; the coating of the quality control line has specific identification Rabbit anti-avidin antibody for avidin. The present invention has the following advantages: the detection line and the quality control line of the present invention adopt independent reaction systems without interference and influence on each other, and the T / C value is used for calibration to ensure the accuracy of test results.

Description

technical field [0001] The invention belongs to the technical field of immunodiagnosis, and in particular relates to an immunochromatographic test strip for quantitatively detecting anti-Müllerian hormone (AMH) by fluorescence and a preparation method thereof. Background technique [0002] Anti-Müllerian hormone (AMH), a member of the transforming growth factor-β (TGF-β) superfamily, is a homodimer consisting of two 55kDa N-terminals and two 12.5kDa C-terminals The constituent glycoproteins are non-covalently bonded disulfide bonds. For males, AMH hormone is produced by testicular Sertoli cells, starting from embryogenesis and throughout life. During embryonic development, AMH can cause the degeneration of Mullerian ducts to form normal male reproductive tracts. In women, AMH hormone is produced by follicular granulosa cells until female menopause, AMH hormone levels gradually decline to undetectable levels. [0003] Studies have shown that: AMH plays an important role in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74
CPCG01N33/74
Inventor 胡鹍辉夏福臻钱纯亘张赛宋永波
Owner SHENZHEN YHLO BIOTECH
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