Polymerase chain reaction detection method of hepatitis B virus genotyping

Abstract

The invention provides a polymerase chain reaction (PCR) detection method of hepatitis B virus (HBV) genotyping. The method comprises the following steps: a primer is designed according to the entire genome sequence of the HBV genotype; in the existence of the primer, DNAs are extracted from a serum sample to perform PCR amplification in a fluorescence PCR meter, the PCR product is analyzed according to the melting curve, the genotype is judged according to the Tm value of the PCR product and the HBV genotyping can be realized. The PCR melting curve method used in the invention is convenient to operate, the detection result of the method has higher coincidence rate with the detection result obtained by adopting a commercial genotyping kit; and the method can be used in the clinical detection of the HBV genotype. On the basis of the PCR technology, the PCR melting curve method which is to judge the HBV genotype according to the Tm value, is provided in the PCR detection method, thus the HBV genotype can be identified conveniently, rapidly and accurately.

Description

technical field [0001] The invention relates to a PCR melting curve detection method for HBV genotyping. Background technique [0002] Hepatitis B virus (HBV) mainly causes liver damage through the host's immune mechanism. The body's immune system recognizes viral antigens and attacks infected liver cells to cause inflammation. This process is affected by multiple factors including the host and the virus. Viral gene heterogeneity affects the expression of antigens and also plays an important role in this process. Different strains have different frequencies of certain mutations, different strains have different resistance to immune clearance of the body, and other factors may lead to different genotypes having different post-infection disease spectrum. [0003] HBV is usually divided into eight genotypes, A to H, based on the heterogeneity of the whole gene sequence ≥ 8%, or the heterogeneity of the S gene sequence ≥ 4%. HBV genotypes were distributed in certain geographic...

AI Technical Summary

Problems solved by technology

There are already some HBV genotyping methods, and the information provided by the sequencing method is comprehensive and reliable, but this method is cumbersome, time-consuming, and has high experimental conditions and requirements, so it is not suitable for widespread development

The results of other methods are not as reliable as the sequencing method, and they are still not standardized and commercialized. For example, PCR-RFLP is cumbersome to operate, and the restriction site is easily affected by gene variation to affect the result judgment; PCR microplate nucleic acid molecular hybridization-ELISA This method is easy to operate and is suitable for general laboratory determination, but it cannot genotype HBV infection with negative serum HBsAg; type-specific primer PCR operation is relatively simple and the result is accurate, but two rounds of PCR and product electrophoresis are still required. , there is a possibility of product contamination

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