A kind of hepatitis E virus IgG antibody colloidal gold method detection kit and preparation method thereof

A technology of hepatitis E virus and detection kit, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long detection time and complicated operation of ELISA method, and achieve the effect of simple operation, low cost and accurate sensitivity

Inactive Publication Date: 2011-12-21
北京中检安泰诊断科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the ELISA method is complicated to operate and takes a long time to detect. It needs to perform operations such as dilution, incubation, separation, washing and color development of the sample to be tested, which takes more than two hours.

Method used

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  • A kind of hepatitis E virus IgG antibody colloidal gold method detection kit and preparation method thereof
  • A kind of hepatitis E virus IgG antibody colloidal gold method detection kit and preparation method thereof
  • A kind of hepatitis E virus IgG antibody colloidal gold method detection kit and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] 1. Source of raw materials

[0069] The source and consumption of each raw material and diluent prepared by Hepatitis E virus IgG antibody detection kit (colloidal gold method) are shown in the following table 10:

[0070] Table 10

[0071]

[0072] 2. Raw material requirements:

[0073] 2.1 Recombinant hepatitis E virus antigen

[0074] (1) Appearance: colorless transparent liquid;

[0075] (2) Concentration and purity requirements: the concentration is greater than 2 mg / ml, determined by SDS-PAGE, and only one band exists under the condition of 10 μl of sample loading;

[0076] 2.2 Goat anti-mouse IgG antibody

[0077] (1) Appearance: colorless transparent liquid;

[0078] (2) Concentration requirements: concentration greater than 4mg / ml;

[0079] 2.3 Mouse anti-human IgG monoclonal antibody

[0080] (1) Appearance: colorless transparent liquid;

[0081] (2) Concentration and purity requirements: the concentration is greater than 2mg / ml, detected by SDS-PAG...

Embodiment 2

[0144] Kit preparation one

[0145] (1) Preparation of reaction membrane: Dilute the recombinant hepatitis E virus antigen to a coating concentration of 1.5 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use The membrane machine draws the two kinds of coating solutions onto the nitrocellulose membrane respectively, and saves the coated nitrocellulose membrane after drying;

[0146] (2) Preparation of mouse anti-human IgG monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgG monoclonal antibody solution was obtained by coupling mouse anti-human IgG monoclonal antibody with a labeling concentration of 10 μg / ml to colloidal gold. Centrifuge at a speed of 12000r / min for 40min, discard the supernatant, add colloidal gold conjugate diluent to 1 / 2 of the original volume, and then use the mixed solution to soak the gold standard pad at 1.5ml / bar to prepare the HEV-IgG gold conjugate pad ,...

Embodiment 3

[0150] Kit Preparation II

[0151] (1) Preparation of reaction membrane: Dilute the recombinant hepatitis E virus antigen to a coating concentration of 1.5 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use The membrane machine draws the two kinds of coating solutions onto the nitrocellulose membrane respectively, and saves the coated nitrocellulose membrane after drying;

[0152] (2) Preparation of mouse anti-human IgG monoclonal antibody gold conjugate pad: Colloidal gold-labeled mouse anti-human IgG monoclonal antibody solution was obtained by coupling mouse anti-human IgG monoclonal antibody with a labeling concentration of 23 μg / ml to colloidal gold. Centrifuge at a speed of 12000r / min for 40min, discard the supernatant, add colloidal gold conjugate diluent to 1 / 2 of the original volume, and then use the mixed solution to soak the gold standard pad at 1.5ml / bar to prepare the HEV-IgG gold conjugate pad , ...

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Abstract

The invention discloses a colloidal gold method detection kit for hepatitis E virus IgG antibody, which comprises recombinant antigen HEV-Ag coated on a nitrocellulose membrane detection line, goat anti-mouse IgG antibody coated on a quality control line and a gold standard Colloidal gold-labeled mouse anti-human IgG monoclonal antibody coated on the pad, the recombinant hepatitis E virus antigen concentration is greater than 2mg / ml, measured by SDS-PAGE; the sheep anti-mouse IgG antibody concentration is greater than 4mg / ml; The concentration of the mouse anti-human IgG monoclonal antibody is greater than 2mg / ml, and it is determined by SDS-PAGE, and the sample amount is 10 μl under the condition of two bands. The invention has the beneficial effects that: the hepatitis E virus IgG antibody detection kit has the characteristics of fastness, convenience, accuracy and high sensitivity, and the whole operation time only needs 20 minutes to interpret the result. Colloidal gold rapid detection test strips, using multi-epitope recombinant antigens as raw materials, have the characteristics of easier operation, low cost, good specificity and high sensitivity, can be detected in a single part, and are easy to popularize. The detection and control effect is obvious.

Description

technical field [0001] The invention relates to an antibody detection kit and a preparation method thereof, in particular to a hepatitis E virus IgG antibody colloidal gold method detection kit and a preparation method thereof. Background technique [0002] Hepatitis E virus (hepatitis E virus, hepatitis E) is the pathogen of non-A non-B acute hepatitis, mainly through the fecal-oral route, and it has also been reported that it can be transmitted through blood. Humans are generally susceptible to hepatitis E, with severe symptoms and high fatality rate. The mortality rate of hepatitis E in young and middle-aged adults can reach 1% to 2%, which is far higher than other types of hepatitis. The mortality rate of hepatitis E in pregnant women can reach as high as 20%. left and right, posing a serious threat to human health. [0003] ELISA is currently the most commonly used method for detecting serum hepatitis E antibodies, among which indirect ELISA is the most widely used, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/531G01N33/532
Inventor 李习荣
Owner 北京中检安泰诊断科技有限公司
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