Primers and method for detecting drug resistance mutation site of hepatitis B virus

A technology of hepatitis B virus and drug-resistant mutation sites, which is applied in the field of medical detection to achieve the effect of large throughput and simple operation

Inactive Publication Date: 2011-09-14
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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Problems solved by technology

This method can detect 5% of the mutant strains in the virus quasi-species pool, but new probes must be designed for emerging mutations, forcing the technology to be updated regularly

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  • Primers and method for detecting drug resistance mutation site of hepatitis B virus
  • Primers and method for detecting drug resistance mutation site of hepatitis B virus
  • Primers and method for detecting drug resistance mutation site of hepatitis B virus

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Embodiment Construction

[0049] The present invention will be further described below in conjunction with embodiment.

[0050] The invention detects the drug resistance mutation site of the hepatitis B virus based on the 454 sequencing technology. First, according to the homology analysis of the nucleotide sequence of the hepatitis B virus genome, 4 pairs of primers were designed, and the 4 PCR products obtained by amplification covered the entire reverse transcriptase gene sequence in an overlapping manner, and the DNA obtained by PCR amplification, etc. The molar ratio was mixed to construct a DNA library of reverse transcriptase gene, and then sequenced by 454 sequencing technology to monitor whether the sample contained drug resistance mutation sites. Compared with the existing detection technology of hepatitis B drug resistance mutation site,

[0051] The present invention will be further described below in conjunction with implementation examples and accompanying drawings.

[0052] 1. Primers ...

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Abstract

The invention discloses primers and a method for detecting a drug resistance mutation site of hepatitis B virus. The invention provides four pairs of PCR (polymerase chain reaction) primers for detecting the drug resistance mutation site of the hepatitis B virus, the four pairs of the PCR primers are used for amplifying DNA (deoxyribonucleic acid) of a genome of the hepatitis B virus, and obtained four fragments are covered on the whole reverse transcriptase gene sequence of the hepatitis B virus in the overlapping way. The four PCR fragments are mixed according to the equal molar ratio for preparing a reverse transcriptase gene bank, 454 high-throughput sequencing and bioinformatics analysis are performed, and whether the hepatitis B virus in a blood serum sample of a patient with the hepatitis B virus contains the drug resistance mutation site or not can be detected. By adopting the primers and the method, above 1% of drug resistance mutation of the hepatitis B virus in blood serum of the patient can be detected, the method has the advantages of high sensitivity, simpleness in operation, large throughput and the like, and the analysis of the drug resistance mutation site of a plurality of samples can be performed by sequencing once.

Description

technical field [0001] The invention relates to the detection of drug-resistant mutation sites of hepatitis B virus, in particular to the detection of drug-resistant mutation sites of hepatitis B virus by using 454 sequencing technology, which belongs to the technical field of medical detection. Background technique [0002] Hepatitis B virus (HBV), referred to as hepatitis B virus, is a DNA virus belonging to the family Hepadnaviridae. According to current knowledge, HBV is only susceptible to human and orangutan, causing HBV disease. The complete hepatitis B virus is in the form of particles with a diameter of 42 nanometers. It was first discovered by British scientist Dane in 1970, so it is also called Dane particles (Dan's particles). Dane particles are composed of outer membrane and inner core. The outer membrane is 7 nanometers thick and is composed of protein and membrane lipid, called hepatitis B virus surface antigen (HBsAg). The diameter of the central part is ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
Inventor 尹飞飞王华林邓菲胡志红
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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