Method for detecting human papilloma virogene type

Abstract

The invention provides a method for detecting the HPV genotypes, with the gene to be detected being one or several types of the type 17 HPV, comprising the following steps: according to the variant sites of the selected HPV genotype universal primer sequence to be detected, an amplification primer aiming at each type is designed; the specific extension primer of each type is designed; (2) PCR amplification is conducted; (3) SAP enzyme treatment is conducted; (4) extension reaction is conducted, among the extending products and the extending primers, the difference of the molecular weight among the extending products of each type is not less than 9D; (5) resin is used to purify extension reaction products; and (6) mass spectrometry detection is conducted, and the type of HPV gene to be detected is determined. By using the method, the invention solves the problems of some of existing detection methods that the typing can not be realized, the multiple infections can not be detected, the accuracy is limited, the throughput is low, the cost is high, and the stability of reaction may be affected as the probe is RNA.

Description

technical field [0001] The invention relates to a method for detecting human papilloma virus genotype. Background technique [0002] HPV is a small non-enveloped DNA virus with a double-stranded closed-loop DNA genome, 7.2-8kb, and a relative molecular weight of 5×10 6 , the HPV genome can be divided into 3 regions: the non-coding upstream regulatory region, the early open reading frame, including E6, E7, E1, E2, E4, E5, the late open reading frame region L1 (the main capsid protein) and L2 (minor capsid protein). According to the difference of the open reading frame (ORF) gene sequence encoding capsid protein L1, the difference of ORF in the L1 region is less than 60%, and it is divided into different genera, and between 60-70%, it is divided into different species. 71-89% are divided into different types. Currently, there are more than 130 types of HPV identified. According to the pathogenicity of the virus, they are divided into high-risk types, such as HPV16, HPV18, HP...

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved by the embodiments of the present invention is to provide a method for detecting human papillomavirus genotype, which aims to solve the problem that existing detection methods cannot be genotyped, cannot detect multiple infections, have limited accuracy, and have low throughput and high cost , and because the probe is RNA, it may affect the stability of the reaction

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