Method and kit for detecting high-risk HPV (human papillomavirus) E6/E7 mRNA (messenger ribonucleic acid) by ligase

Abstract

The invention relates to a method and a kit for detecting high-risk HPV (human papillomavirus) E6 / E7 mRNA (messenger ribonucleic acid) by ligase. The method is characterized in that specific connection primers are connected into a cDNA (complementary deoxyribonucleic acid) fragment complementary with a target sequence of the HPV E6 / E7 mRNA under the action of the ligase, and the successfully connected cDNA fragment can be amplified by a pair of universal primers in a later fluorescent PCR (polymerase chain reaction) system, in other words, eighteen high-risk HPVs capable of causing cervical cancer can be detected in the same amplification system by the single fluorescent PCR technology, so that the problems, such as low amplification efficiency and sensitiveness, caused by excessive primers are solved effectively. The aim of HPV E6 / E7 mRNA detection is to screen out high-risk groups in HPV-infected people groups, so that the method has more significance in clinical diagnosis as compared with existing HPV genome L1 distinguishing type detection methods.

Description

technical field [0001] The invention relates to a high-risk HPV fluorescence detection kit, in particular to a method and kit for detecting high-risk HPV VE6 / E7 mRNA by using ligase. Background technique [0002] Cervical cancer is the most common gynecological malignancy. The high-incidence age of carcinoma in situ is 30-35 years old, and that of invasive carcinoma is 45-55 years old. In recent years, the onset tends to be younger. Early detection and treatment of cervical cancer is the key to effective treatment of cervical cancer. [0003] Existing studies have shown that the main cause of cervical cancer is persistent or recurrent infection of high-risk human papillomavirus (HPV). HPV testing has become an important part of cervical cancer screening and health management in many countries around the world. Human papillomavirus belongs to the papillomavirus genus of the Papillomaviridae family, and is a spherical non-enveloped double-stranded DNA virus with a diameter ...

AI Technical Summary

Problems solved by technology

[0005] The disadvantages of the existing techniques for genotyping detection of high-risk human papillomavirus by fluorescent PCR method are as follows: ① Generally, the L1 gene of the HPV genome is detected, and the detection results are not as clinically diagnostic as the detection results of the E6 / E7 genes

②In the early stage, cumbersome HPV nucleic acid extraction is generally required; ③After the nucleic acid extraction is completed, multiple PCR amplification is performed using the common Taqman probe method, which means that a reaction tube contains multiple primers, and the mutual interference between the primers leads to amplification sensitivity. Low and time-consuming; ④Generally, FAM and HEX are two fluorescent channels for detection, which cannot specifically distinguish high-risk HPV types, and it is difficult to meet the needs of actual use

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