Primer and typing probe combination for detecting human papilloma virus and application of primer and typing probe combination

Abstract

The invention provides a primer and typing probe combination for detecting human papilloma virus and application of the primer and typing probe combination. The primer and typing probe combination for detecting the human papilloma virus comprises a universal primer pair and a typing probe, wherein the universal primer pair is used for amplifying a human papilloma virus genome advanced region L1; the universal primer pair comprises an upstream primer group and a downstream primer group; the upstream primer group comprises an upstream primer 1, an upstream primer 2 and an upstream primer 3; the upstream primer group comprises nucleotide sequences as shown in SEQ ID No. 1 to SEQ ID No. 3; the downstream primer group comprises a downstream primer 1, a downstream primer 2 and a downstream primer 3; and the downstream primer group comprises nucleotide sequences as shown in SEQ ID No. 4 to SEQ ID No. 6. The primer and typing probe combination for detecting the human papilloma virus can detect and distinguish 28 HPV subtypes, is superior to similar products in China at present, and is wider in coverage and more representative.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a combination of primers and typing probes for detecting human papillomaviruses and an application thereof. Background technique [0002] Cervical cancer is the most common gynecological malignancy, and its incidence tends to be younger. Early screening, discovery and treatment of cervical cancer are the key to effective prevention and treatment of cervical cancer. Studies have shown that human papillomavirus (Human Papillomavirus, HPV) infection is directly related to the occurrence of cervical cancer. HPV belongs to the papillomaviridae family, is a small molecule, non-encapsulated circular double-stranded DNA virus, which can cause the proliferation of squamous epithelium of human skin and mucous membranes. At present, there are more than 190 HPV subtypes identified, about 50 of which infect the stratified squamous epithelium of human genital tract skin and mucous mem...

AI Technical Summary

Problems solved by technology

[0008] Real-time fluorescent PCR technology still has unavoidable defects, including: it can only detect whether the sample is infected with the corresponding genotype of HPV, but it does not have the specific typing function, the detection of a single subtype takes a long time, and multiple subtypes can be detected at the same time. During detection, the interference between primers will lead to a decrease in detection sensitivity; the early stage of real-time fluorescent PCR generally needs to go through a cumbersome nucleic acid extraction process; a relatively expensive fluorescent quantitative PCR instrument is required

[0009] HC-2 technology also has inherent defects, including: low detection specificity, about 85%, slightly lower than liquid-based cytology; HC-2 technology belongs to the comprehensive application of nucleic acid hybridization, immune reaction and chemiluminescence technology, detection The complexity and cost are high; HPV genotyping cannot be performed accurately; the stability of RNA probes is poor, and the requirements for experimental conditions and operating techniques are also high; it is easy to cause cross-reactions and lead to false positive results, and high-risk types There is a certain cross-reaction with the low-risk type

[0010] The gene chip RDB technology also has its defects, including: multiple sets of primers for different genotypes of HPV need to be designed in the early stage, the PCR reaction system is relatively complicated, and the detection sensitivity is reduced or even missed due to interference between primers in actual amplification. inspection; due to the use of many primers, the corresponding product production process is complicated, which greatly increases the cost

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