High-throughput sequencing detection method used for HPV typing and integration

Abstract

The invention discloses a high-throughput sequencing detection method used for HPV typing and integration. According to the method, the genes of current HPV subtypes are selected, in combination witha second-generation high-throughput sequencing technology, the type of HPV infected by a patient is detected more comprehensively, and the method overcomes the difficulties that a traditional detection method is low in accuracy rate, high in false positive result, low in repeatability and high in rate of missed diagnosis. In the field of molecular diagnosis, the mostly direct and specific technology is gene sequencing, and the second-generation high-throughput sequencing technology has the advantages of higher detection flux, higher sequencing speed, higher accuracy, lower cost and more abundant information contents compared with a classical Sanger sequencing method mostly adopted at present. According to the method, with the help of the second-generation high-throughput sequencing technology, accurate typing can be carried out on high-risk HPV and low-risk HPV, whether the integration of a human genome occurs is detected, accurate individual assessment is carried out on a detector, and the risk of a disease is prevented, so that the occurrence of a tumor is prevented.

Description

technical field [0001] The invention relates to a high-throughput sequencing detection method for HPV typing and integration. Background technique [0002] Human papillomavirus (HPV) is a small DNA virus that is highly specific for mucous membranes and epithelial skin. HPV virus is mainly composed of DNA core and protein capsid. Its genome is a double-stranded circular DNA, about 7.5-8.0 kb in length, containing 8 open reading frames (ORFs), divided into 3 regions according to different functions: (1) early region (early region, E region) : Encode six early proteins including E1, E2, E4, E5, E6, and E7 respectively, which are involved in viral DNA replication, transcription, translation regulation, and transformation; (2) late region (L region): encodes the main Coat protein L1 and minor coat protein L2; (3) non-coding region (uncoding region, UCR), also known as long control region (LCR) or upstream regulatory region (URR): contains the replication origin of HPV genomic D...

AI Technical Summary

Problems solved by technology

Laboratory technicians will observe abnormal cells under a microscope to diagnose the patient's condition. However, since this method only uses observation as the basis for diagnosis, its accuracy is low and false due to the influence of material collection, smear production quality, and film reading technology. High negative, poor repeatability, the result and analysis missed diagnosis rate can reach 30%

Among the genotyping methods for checking and screening HPV, real-time fluorescent PCR method and hybridization capture method are the most commonly used methods, but the diagnostic operation for different genotypes of HPV is cumbersome, and the detection throughput is low, which is difficult to meet the requirements of clinical simultaneous detection of multiple subtypes. Type needs, not suitable for large-scale screening of cervical cancer

Commercial hybrid capture kits can detect HPV DNA in clinical samples without PCR amplification, and can distinguish between high-risk and low-risk types, but such kits cannot identify the specific genotypes and Whether it is integrated or not, there are more false negatives and false positives, and the sensitivity and specificity need to be improved

However, in the field of molecular diagnosis, the most direct and clear technology is gene sequencing. The traditional gene sequencing method is carried out through the Sanger termination method. This method can only read the sequence of a single gene at a time. disease, it is difficult to obtain ideal sequencing results

These existing detection methods can only perform a small amount of HPV typing, but cannot detect whether HPV is integrated into the human genome, and whether HPV is integrated into the human genome is a prerequisite for HPV to cause cancer

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