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Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone

A technique for quantitative detection of Müllerian hormones, applied in the biological field, can solve the problems of expensive chemiluminescence and electrochemiluminescence equipment, unsuitable for single person and small batch detection, long detection time, etc., and can shorten the detection time. Detection time, suitable for professional department operation, easy operation effect

Inactive Publication Date: 2016-08-24
付国亮
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ELISA method is widely used in hospital laboratories due to its accurate quantification, but the detection time is long and inconvenient; chemiluminescence and electrochemiluminescence methods are currently used in clinical applications due to expensive equipment and are not suitable for single-person and small-batch detection. still relatively few

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  • Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone
  • Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone
  • Test strip for quantitatively detecting anti-mullerian hormone, preparation method thereof and determination method for concentration of anti-mullerian hormone

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preparation example Construction

[0039] The preparation method of the above-mentioned test strip comprises the following steps: sequentially fixing a sample absorbent pad, a labeled antibody pad, a coating film and a water-absorbent pad on the bottom plate of the test strip;

[0040] Described coated film is prepared by following method:

[0041] Use the first phosphate buffer to dilute the rabbit anti-mouse IgG and AMH antibody respectively to obtain the rabbit anti-mouse IgG solution and the AMH antibody solution, the concentration of the first phosphate buffer is 0.01-0.03M, and the pH value is 7.2- 7.6, the concentrations of the rabbit anti-mouse IgG solution and the AMH antibody solution are both 1mg / ml-1.5mg / ml; use a quantitative spray film instrument to mix the two at 0.5cm-1.0μl / cm Spray on the nitrocellulose membrane at an interval of cm, dry at 35-38°C for 0.5-1.5h, add a desiccant and seal it for later use.

[0042] The labeled antibody pad is prepared by the following method:

[0043] Preparati...

Embodiment 1

[0048] Preparation of fluorescent immunochromatographic test strips for AMH:

[0049] A. Antibody preparation: select commercial AMH monoclonal antibody, dialyze overnight (18h) at 4°C with 0.05M pH8.5 borate buffer solution; select commercial AMH monoclonal antibody, use 0.02M pH7.4 Dialysis against PBS at 4°C overnight (16h);

[0050] B, the preparation of the fluorescent microsphere cushion of labeling AMH monoclonal antibody: select the fluorescent microsphere (Bangslab company, excitation wavelength 350nm, detection wavelength 615nm) that diameter is 100nm for use, with 0.05M, pH4.5MES damping fluid adjusts microsphere concentration to be The mass fraction was 1%, and then the AMH monoclonal antibody was labeled on the fluorescent microspheres by covalent coupling with carbodiimide (EDC) and succinimide (NHS). Spray the prepared fluorescent microspheres on the fluorescent microsphere pad at an amount of 3 μl / cm using a quantitative film sprayer;

[0051] C. Preparation ...

Embodiment 2

[0060] The test strip preparation method of the present embodiment comprises the following steps:

[0061] A. Antibody preparation: select commercial AMH monoclonal antibody, dialyze overnight at 4°C with 0.05M pH8.5 borate buffer; select commercial AMH monoclonal antibody, use 0.02M pH7.4 PBS 4 Dialysis at ℃ overnight;

[0062] B. Preparation of colloidal gold pads labeled with AMH monoclonal antibodies:

[0063] Take 50ml of colloidal gold and filter.

[0064] Dilute the antibody 3 times with water (about 1 mg / ml), add it within 30 minutes, and mix well while adding it. The mixing force should be small near the ice to prevent damage to the connection between the antibody and the colloidal gold.

[0065] 10% BSA was added to make the final concentration of the solution 1%, and 10% PEG was added to make the final concentration 0.2%, and the addition was completed within 15 minutes.

[0066] Centrifuge at 10000r / min for 60min, carefully suck out the supernatant, and resuspen...

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Abstract

The invention discloses a test strip for quantitatively detecting anti-mullerian hormone, a preparation method thereof and a determination method for the concentration of the anti-mullerian hormone, and belongs to the technical field of biology. The test strip comprises a base plate. A sample absorption pad, a labeled antibody pad, a coating film and a water absorption pad are sequentially attached to the base plate. The test strip has the advantages of being accurate in detection result, convenient to use and low in cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an immunochromatographic test strip for quantitative detection of human anti-Müllerian hormone (AMH) and a preparation method thereof and an instant and rapid detection of the concentration of anti-Mullerian hormone (AMH) test methods. Background technique [0002] With the change of modern life style, the general phenomenon of late marriage and late childbearing, unhealthy modern life style (such as excessive dieting, smoking and drinking, staying up late), and huge work pressure have caused the rate of infertility to rise rapidly. Therefore, early assessment of female fertility is very necessary. Anti-Müllerian hormone is such an innovative evaluation indicator of ovarian reserve function; anti-Mullerian hormone, also known as AMH, is the direct product of early follicles, a sugar composed of two 72KDa monomers connected by a disulfide bridge Protein dimer, secreted by granulosa ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/558G01N33/532G01N33/533
CPCG01N33/532G01N33/533G01N33/558G01N33/577
Inventor 付国亮段学军
Owner 付国亮
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