Probe and detection method for HPV (Human Papillomavirus) detection

A probe and reaction technology, applied in the field of genetic engineering, can solve the problems of low efficiency and high cost

Inactive Publication Date: 2019-08-27
杭州链康医学检验实验室有限公司
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  • Abstract
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  • Application Information

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Problems solved by technology

Although this method has good specificity and high sensiti...

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  • Probe and detection method for HPV (Human Papillomavirus) detection
  • Probe and detection method for HPV (Human Papillomavirus) detection
  • Probe and detection method for HPV (Human Papillomavirus) detection

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Embodiment approach

[0062] General implementation scheme: a kind of probe and detection method for HPV detection comprise the following steps:

[0063] (1) Prepare the probe;

[0064] (2) Prepare the DNA of cervical exfoliated cell samples with complete electrophoresis main band and no obvious degradation;

[0065] (3) Fragmentation of genomic DNA using ultrasonic physical fragmentation method;

[0066] (4) Purifying the fragmented product in step (3) with LC magnetic beads;

[0067] (5) constructing the fragmented DNA into an Illumina genome library;

[0068] (6) Amplify the genomic library by PCR reaction and use LC magnetic beads to purify the amplified product;

[0069] (7) Hybridizing the probe prepared in step (1) with the genomic library in step (6);

[0070] (8) capture the hybridization product with streptavidin magnetic beads and wash the non-specific binding;

[0071] (9) PCR amplification of target DNA fragments bound to the streptavidin magnetic beads;

[0072] (10) High-throug...

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Abstract

The invention discloses a probe and a detection method for HPV (Human Papillomavirus) detection. The preparation process of the probe requires two rounds of PCR reactions. The NGS technology-based detection method comprises the steps of preparing a DNA template, performing fragmentation on genomic DNA, purifying a fragmented product, constructing a genomic library, performing PCR amplification onthe genomic library, hybridizing the probe with the genomic library, capturing a hybridization product by magnetic beads, and performing high-throughput sequencing on the captured product and analyzing whether a sample to be tested is infected with HPV, the infection degree of the HPV, the integration location and the pathogenic mechanism of the HPV and the like. The probe and the method disclosedby the invention have the advantages of low cost, fast data analysis, high coverage depth, high detection accuracy and the like, and are suitable for early screening and postoperative diagnosis of diseases caused by the HPV, and provide guidance for personalized medication.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, and more specifically relates to a probe and a detection method for HPV detection. Background technique [0002] Human papillomavirus (Human Papillomavirus, HPV) is an epitheliophilic virus with high specificity. It is a small DNA virus with double-stranded closed circle and contains about 8000 base pairs. HPV can be integrated into the host cell genome to cause instability of the host cell genome, resulting in chromosomal inversion, translocation, deletion, and rearrangement, which makes the genome structure of the host cell abnormal, and can cause insertional mutagenesis leading to key tumor suppressor genes of the host cell. Abnormal inactivation or abnormal activation of oncogenes, and virus integration can also cause copy number variation in certain regions of the host cell genome, indirectly affecting the expression levels of some key oncogenes or tumor suppressor genes in host ...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6806C12N15/11C12Q1/6869C12R1/93
CPCC12Q1/708C12Q1/6806C12Q1/6869C12Q2531/113C12Q2563/143C12Q2563/149C12Q2535/122
Inventor 郎秋蕾钱刚方超蔡霖霖殷楠楠刘歆李璐璐
Owner 杭州链康医学检验实验室有限公司
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