A colloidal gold method detection kit for hepatitis A virus igm antibody and preparation method thereof

A technology of hepatitis A virus and detection kit, which is applied in the direction of measuring devices, diseases resistant to vector transmission, instruments, etc., can solve the problems of long detection time and complicated operation of ELISA method, and achieve simple operation, low cost and obvious effect Effect

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 colloidal gold method detection kit for hepatitis A virus igm antibody and preparation method thereof
  • A colloidal gold method detection kit for hepatitis A virus igm antibody and preparation method thereof
  • A colloidal gold method detection kit for hepatitis A virus igm antibody and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] 1. Source of raw materials

[0072] The source and dosage of each raw material and diluent prepared by the Hepatitis A virus IgM antibody detection kit (colloidal gold method) are shown in the following table 10:

[0073] Table 10. Sources and dosages of raw materials and diluents

[0074]

[0075]

[0076] 2. Raw material requirements:

[0077] 2.1 Recombinant HAV antigen

[0078] (1) Appearance: colorless transparent liquid;

[0079] (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;

[0080] 2.2 Goat anti-mouse IgG antibody

[0081] (1) Appearance: colorless transparent liquid;

[0082] (2) Concentration requirements: concentration greater than 4mg / ml;

[0083] 2.3 Mouse anti-human IgM monoclonal antibody

[0084] (1) Appearance: colorless transparent liquid;

[0085] (2) Concentration and purity requirements: the concentra...

Embodiment 2

[0144] Kit preparation one

[0145] (1) Preparation of reaction membrane: Dilute the recombinant HAV antigen to a coating concentration of 1.0 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use a film dispenser to The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;

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

Embodiment 3

[0150] Kit Preparation II

[0151] (1) Preparation of reaction membrane: Dilute the recombinant HAV antigen to a coating concentration of 1.0 mg / ml with phosphate buffer, dilute the goat anti-mouse IgG antibody to a coating concentration of 2.0 mg / ml, and use a film dispenser to The two coating solutions were drawn onto the nitrocellulose membrane respectively, and the coated nitrocellulose membrane was dried and stored;

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

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Abstract

The invention discloses a colloidal gold detection kit for hepatitis A virus IgM antibody, comprising a recombinant antigen HAV-Ag coated on a nitrocellulose membrane detection line, a goat anti-mouse IgG antibody and a gold label coated on a quality control line The mouse anti-human IgM monoclonal antibody labeled with colloidal gold coated on the pad, the recombinant hepatitis A virus antigen concentration was greater than 2mg / ml, and was determined by SDS-PAGE; the goat anti-mouse IgG antibody concentration was greater than 4mg / ml; The concentration of the mouse anti-human IgM monoclonal antibody was greater than 2 mg / ml, which was determined by SDS-PAGE, and the loading amount was two bands under the condition of 10 μl. The beneficial effects of the invention are as follows: the hepatitis A virus IgM antibody detection kit has the characteristics of rapidity, simplicity, accuracy and high sensitivity, and the whole operation time only takes 20 minutes to interpret the results. The colloidal gold rapid detection test strip is based on the multi-epitope recombinant antigen as the raw material. It has the characteristics of easier operation, low cost, good specificity and high sensitivity. It can be detected in a single batch and is 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 A virus IgM antibody colloidal gold method detection kit and a preparation method thereof. Background technique [0002] The pathogen of hepatitis A (hepatitis A, referred to as HA) is hepatitis A virus (HAV), which is a small RNA virus (picornavirus) with a diameter of 27 nm. The virus exists in the patient's feces, serum, bile, and liver cytoplasm. It has strong resistance in vitro and can survive being heated at 60°C for 4 hours; it can tolerate the environment of pH 3.0; it can survive for a long time at low temperature. [0003] The commonly used detection methods for hepatitis A serum antibodies mainly include hemagglutination test, enzyme-linked immunoassay, and radioimmunoassay. One of the most extensive laboratory testing methods. Anti-HAV-IgM is the main indicator for early diagnosis of hepatitis A. The appearance of serum IgM ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/576G01N33/531G01N33/532
CPCY02A50/30
Inventor 李习荣
Owner 北京中检安泰诊断科技有限公司
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