Detection method of hepatitis A viral antigen

Abstract

The invention provides a detection method of hepatitis A viral antigen. The method comprises the steps: long arm biotin is adopted to mark HAV-IgG, horse radish peroxidase is adopted to mark streptavidin to be used as an amplifying system, when the hepatitis A viral antigen is detected, as the long arm biotin has longer space arm than common biotin, the sterically hindered can be reduced, and a plurality of biotin molecules can be simultaneously combined with an avidin compound; moreover, as water-soluble biotin, the long arm biotin marks the hepatitis A viral antigen directly, can reduce the steps of the immunological detection and the detection time, namely the total detection time can be reduced to 3.5 hours from usual 2 days, the background can be reduced through reducing cross reaction, and the detection sensitivity can be improved; in addition, as the streptavidin marked by the horse radish peroxidase is different from avidin, the molecular weight is about 60000, carbohydrate is not included, and the nonspecific binding is lower than the avidin, therefore the method has the effects of improving sensibility and specificity.

Description

technical field [0001] The invention relates to a detection method of hepatitis A virus (HAV) antigen, belonging to the technical field of biology. Background technique [0002] In the research work of hepatitis A live attenuated vaccine, hepatitis A inactivated vaccine and related viruses, detection of hepatitis A virus antigen is required, and because hepatitis A virus HAV cannot produce cytopathic changes (CPE), so In the detection of virus infectivity titer, after the virus to be tested is multiplied on the cells, the virus must be released by physical or chemical methods, and then the hepatitis A virus antigen (HAAg) detection is carried out by related methods. At present, there is no special hepatitis A virus antigen detection kit for sale on the market, and the detection of antigens is performed by various manufacturers or scientific research institutions according to their own needs. According to literature search, the current hepatitis A virus antigen detection met...

AI Technical Summary

Problems solved by technology

[0003] Although the detection sensitivity of RT-PCR and RT-PCR-ELISA methods are good, they require special instruments and equipment, the detection cost is high, and the specificity is affected by many factors; the double antibody sandwich method (ELISA) is easy to operate, but it needs to use Different strains of monoclonal antibodies are coated and labeled respectively, and the screening and preparation of monoclonal antibodies is not easy, and it is difficult to establish in ordinary laboratories; the indirect method ( ELISA), is not a specific detection method for hepatitis A virus antigen, the sensitivity is not high, the detection time is relatively long, the detection steps are many, and there are many influencing factors.

However, the reported biotin-avidin-biotinylated horseradish peroxidase method uses biotin to label antibodies, but it uses ordinary fat-soluble biotin, which needs to be dissolved in an organic solvent before it can be used. Labeling, and it uses avidin-bound biotinylated horseradish enzyme complex as the enzyme amplification system, the concentration ratio of the complex is troublesome, there are many steps, and the detection time is long

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