In vitro diagnosis kit of hepatitis B virus E antibody through dual-antigen sandwich method and preparation method thereof

Abstract

The invention provides an in vitro diagnosis kit (an ELISA method) of a hepatitis B virus E antibody through a dual-antigen sandwich method, which comprises the following components: 1) a recombinant HBeAg coated in a solid phase carrier; 2) an enzyme-labeled recombinant HBeAg; and 3) a substrate solution, for example, a chromogenic substrate solution applied to the ELISA, or an enzymatic chemiluminescent substrate solution. Besides, the invention provides a method for preparing the kit, which comprises the following steps: 1) coating the recombinant HBeAg into the solid phase carrier; 2) performing enzyme labeling on the recombinant HBeAg; 3) preparing the substrate solution; and 4) assembling the components into a finished kit. The kit has the advantages of high sensitivity, good specificity, accurate result and strong clinical applicability, and can accurately reflect the amount of the E antibody in the body of a hepatitis B patient.

Description

technical field [0001] The invention relates to the field of immunomedicine analysis, in particular, the invention relates to a hepatitis B virus E antibody in vitro diagnostic kit and a preparation method. Background technique [0002] Hepatitis B Virus belongs to the family of hepatotropic viruses, which can cause acute hepatitis, chronic hepatitis and liver cirrhosis, and is closely related to the occurrence of liver cancer. It is one of the most widespread and deadly viruses in the world today. According to statistics, more than 3.5 billion people in the world are carriers or patients of hepatitis B [Zucherman J.N., et al., J. Infect, 2000, 41(2): 130-136]. China is a high-incidence and endemic area of ​​hepatitis B, with about 120 million hepatitis B patients [Edited by Zhang Jianying, Research Progress in Viral Liver Diseases, First Edition, China Science and Technology Press, 1992]. Moreover, about 1 million people worldwide die each year due to HBV infection [Parki...

AI Technical Summary

Problems solved by technology

Fourth, the color difference between negative and positive is obvious, and it is almost possible to judge the negative and positive results by visual inspection.

When the sodium periodate method is used to couple HBeAg to the enzyme, the amino group within the range of the determinant of the enzyme and HBeAg is more likely to bind to the enzyme

When the enzyme molecule is combined with HBeAg, on the one hand, when the enzyme binds to the amino group on the HBeAg epitope, the epitope will be inactivated; on the other hand, if HBeAg is coupled to more than one enzyme molecule, due to the enzyme The molecular weight is 43,000 Daltons, which is much larger than HBeAg, resulting in steric hindrance, resulting in most of the inactivation of HBeAg

Therefore, if an inappropriate coupling method is used to couple HBeAg to the enzyme, it will cause the inactivation of HBeAg, and the apparent reflection seems to be that HBeAg cannot be coupled to the enzyme.

[0012] In addition, since the recombinant HBeAg prepared today is mixed with the activity of HBcAg, when the enzyme molecule is coupled with HBeAg, the activity of HBcAg still exists, and it will still produce an immune reaction with the hepatitis B core antibody adsorbed on the solid phase, resulting in Cross-reaction to HBV core antibody positive samples

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