Differential detection method and kit for eight genotypes of hepatitis B virus

Abstract

The invention discloses a method and a kit for identifying and detecting eight gene subtypes of hepatitis B virus. Eight gene subtypes A-H of hepatitis B virus are rapidly qualitatively and quantitatively identified and detected by combining multiplex polymerase chain reaction (PCR) with a liquid chip, a primer pair with extremely high sensitivity on the respectively amplified virus specific sequences is selected, a probe with reasonable base composition is designed, and the detection sensitivity is improved. According to the technical scheme in the invention, the eight gene subtypes A-H of hepatitis B virus can be identified in a single test, and the method has the advantages of high flux, high specificity and sensitivity, stable result, high repeatability, simple operation and high detection speed.

Description

technical field [0001] The invention belongs to the technical field of biochips and diagnostic reagents, and relates to a method for identifying and detecting eight subtypes of hepatitis B virus, as well as primers and probes, in particular to a microsphere-based liquid phase chip technology for rapid A method for the detection of eight genotypes of hepatitis B virus. Background technique [0002] Hepatitis B virus is a long-term problem that has plagued the global medical community. It is one of the important pathogenic factors of acute and chronic hepatitis, liver cirrhosis, and liver cancer, and it is also one of the most serious public health problems in the world. According to the WTO report, among the more than 6 billion people in the world, about 3 billion people live in areas with high prevalence of hepatitis B virus (HBV), and more than 2 billion people have been infected with HBV, of which more than 350 million are chronically infected. The annual incidence of hep...

AI Technical Summary

Problems solved by technology

However, these methods have more or less defects: for example, although the results of gene sequencing and typing methods are accurate and reliable, the process is cumbersome, time-consuming, costly, and difficult to promote. It is not suitable for the detection of large samples and lacks sensitivity. Mixed infection of two or more HBV genotypes

The restriction endonuclease cutting site change caused by the polymorphism of a single nucleotide site by PCR-RFLP hinders endonuclease digestion, complex bands will appear if the digestion is not complete, and the recognition of the restriction map is complicated , thus affecting the reliability of genotyping, and this method also cannot find the mixed infection of several genotypes

The nested PCR method requires the synthesis of multiple primers and two rounds of PCR, which takes longer than the RFLP method and has a greater chance of contamination, and it cannot completely avoid the occurrence of non-specific amplification bands that hinder the observation of typing results

INNO-LiPA has the disadvantages of high price and low specificity

Monoclonal antibody ELISA method cannot genotype serum HBsAg-negative HBV infection, and a small number of HBsAg cannot be typed

Gene chips have disadvantages such as high cost, high false positive rate, and insufficient integration of different chips.

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