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HPV typing detection method

A hybridization probe and nucleic acid chip technology, applied in the field of biomedicine, can solve the problems of no accurate and effective HPV typing detection method, and achieve the effect of simple operation, low quality requirements and high sensitivity

Inactive Publication Date: 2016-08-17
SHENZHEN HUAZHONG BIOLOGICAL PHARMA MACHINERY
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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for HPV typing detection, aiming to solve the problem that there is no accurate and effective HPV typing detection method at present

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[0015] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention clearer, the present invention will be further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0016] The embodiment of the present invention provides a method for HPV typing detection, comprising the following steps:

[0017] S01. Provide an in vitro sample containing HPV, use a nucleic acid precipitant to extract nucleic acid, and design biotin-labeled primers and hybridization probes of different types according to the L1 gene target sequence of the HPV;

[0018] S02. Using the primers to perform PCR amplification on the nucleic acid to obtain a PCR product having a nucleic acid fragment of the L1 region of the HPV;

[0019] S03. Immobilizing the hybridization probes of different...

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Abstract

The invention provides an HPV typing detection method. The HPV typing detection method includes the following steps that an in-vitro sample to be detected and containing HPV is provided, and nucleic acid precipitant is adopted to extract nucleic acid according to different types of primers and hybridization probes marked by biotin and designed by L1 gene target sequence of HPV; the primers are used for PCR amplification of nucleic acid to obtain a PCR product with a nucleic acid fragment of an L1 region of HPV; the different types of hybridization probes are fixed to different positions of a hybridization chip and numbered to obtain a hybridization chip with the hybridization probes fixed; the PCR product and the hybridization chip with the hybridization probes fixed are subjected to reverse dot blot hybridization to obtain a nucleic acid chip; the nucleic acid chip is processed with streptavidin marked by alkaline phosphatase and then reacts with a chromogenic substrate BCIP / NBT for color development, and the type of HPV in the in-vitro sample to be detected is judged according to the chromogenic position of the nucleic acid chip.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for HPV type detection. Background technique [0002] At present, domestic HPV (human papillomavirus) detection is mainly divided into HPV antigen detection, HPV antibody detection and HPV-DNA detection. Among them, the detection of HPV antigen: when HPV infects human epidermal cells, it will proliferate in the cells and synthesize capsid proteins. The capsid protein will become the HPV antigenic component. Immunoenzyme staining can detect the HPV antigen components in infected tissue cells, so as to know whether there is HPV infection. The positive detection of HPV antigen will have important reference value for the evaluation of HPV infection or genital warts. But this detection method also has its shortcomings. Only by means of this assay can only confirm the existence of HPV particles, but cannot detect the subclinical and latent types of HPV infe...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/6837C12Q1/701C12Q2563/131C12Q2563/125
Inventor 李立家王璞胡焰文欢肖承荣庄学军王瑢刘舜
Owner SHENZHEN HUAZHONG BIOLOGICAL PHARMA MACHINERY
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