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Human anti-rabies virus IgG antibody ELISA test kit

A detection kit and rabies virus technology, applied in the field of human anti-rabies virus IgG antibody ELISA detection kits, can solve the problems of difficult refolding of recombinant proteins, radioactive isotope contamination, lack of spatial conformational epitopes, etc.

Active Publication Date: 2013-04-10
WUHAN CHOPPER BIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The recombinant protein has the disadvantages of difficult renaturation, lack of spatial conformational epitopes, and the disadvantage of difficult purification of the whole virus, which will lead to low detection sensitivity
Fluorescence immunoassay, radioimmunoassay and chemiluminescence method require certain experimental instruments, and radioimmunoassay still has the problem of radioisotope contamination
Although colloidal gold immunochromatography has the advantage of simple operation, its sensitivity is relatively low.

Method used

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  • Human anti-rabies virus IgG antibody ELISA test kit

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Embodiment Construction

[0014] One. The preparation method of human anti-rabies virus IgG antibody ELISA detection kit

[0015] 1. Preparation of rabies virus monoclonal antibody: immunize BALB / c mice with purified rabies virus antigen to obtain antigen-stimulated spleen cells, fuse the spleen cells with mouse myeloma cell lines, and screen using HAT selective medium Hybridoma cells, use ELISA method and IFA method to identify hybridoma cell lines resistant to rabies virus, and obtain cell lines secreting highly specific monoclonal antibodies after three times of subcloning, and produce monoclonal anti-rabies virus through mouse ascites Antibody;

[0016] 2. Preparation of HRP-mouse anti-human IgG enzyme conjugate: extract and purify human IgG according to conventional methods; immunize BALB / c mice to obtain antigen-stimulated spleen cells, and fuse the spleen cells with the mouse myeloma cell line, Use HAT selective medium to screen hybridoma cells, use ELISA method to identify hybridoma cell lines...

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Abstract

The invention relates to a human anti-rabies virus IgG antibody ELISA test kit. An ELISA plate is firstly coated with an anti-rabies virus monoclonal antibody, wherein the coating buffer solution is a 0.05M carbonate buffer solution of which the pH value is 9.6, and the coating amount is 0.1-1ug per hole; a blocking solution is a BSA or skimmed milk of which the mass concentration is 1-10%; the ELISA plate is coated with a rabies virus purified antigen after being blocked, wherein the coating amount is 0.1-1ug per hole; a sample diluent is a 0.01mol / L phosphate buffer solution (PBS) which contains bovine serum albumin (BSA) with a mass concentration of 0.1-10% and NaN3 with a mass concentration of 0.01-0.05 and has a pH value of 7.2-7.4; an enzyme conjugate is a horse radish peroxidase-mouse anti-human IgG enzyme conjugate; a concentrated cleaning solution is a 0.01mol / L PBS which contains tween-20 with a volume concentration of 0.05% and has a pH value of 7.2-7.4; a zymolyte A solution is a 3,3'-5,5'-tetramethyl benzidine solution, and a zymolyte B solution is an oxydol solution; and a stop solution is a 1mol / L H2SO4 solution, and a positive control and a negative control are arranged in the kit. The specificity of the kit is up to 100%, and the sensitivity is 1:640. The kit is used for evaluating the immunity effect of humans inoculated with rabies vaccines.

Description

technical field [0001] The invention relates to a detection kit, in particular to a human anti-rabies virus IgG antibody ELISA detection kit. Background technique [0002] Rabies is a zoonotic infectious disease caused by Rabies virus (RV), and its main hosts are dogs, cats and other animals. The typical symptom is hydrophobia, also known as: hydrophobia. The disease is extremely dangerous, and the case fatality rate is almost 100%. With the development of the economy and the improvement of people's living standards, the number of pet dogs is not only increasing, but also the area of ​​activity of dogs is constantly expanding. According to the information obtained from the pet medical market, people are eager to know whether pet dogs can gain protection after being vaccinated against rabies. However, the current detection of rabies virus antibody has not been widely used due to the limitation of detection conditions and costs. Rabies virus is the pathogen of rabies, and m...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/543G01N33/535C07K14/145
Inventor 刘汉平廖园园刘洁王威彭杏漆世华温文生谢红玲
Owner WUHAN CHOPPER BIOLOGY
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