Nucleic acid sequence for typing detection of cutaneous human papillomavirus and application thereof

A human papillomavirus and nucleic acid sequence technology, which is applied in the field of skin-type human papillomavirus typing and detection, and can solve the problem of lack of nucleic acid sequences for skin-type HPV detection methods.

Active Publication Date: 2017-06-13
THE FIRST HOSPITAL OF CHINA MEDICIAL UNIV +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the current HPV detection methods are aimed at mucosal HPV, and there is a lack of a mature skin-type HPV detection method and nucleic acid sequence that can be applied clinically.

Method used

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  • Nucleic acid sequence for typing detection of cutaneous human papillomavirus and application thereof
  • Nucleic acid sequence for typing detection of cutaneous human papillomavirus and application thereof
  • Nucleic acid sequence for typing detection of cutaneous human papillomavirus and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Design and screen primer nucleic acid sequences: First, compare the L1 segments of each skin type HPV, select the parts with large differences to design primer nucleic acid sequences, and design multiple pairs of candidate primers for each type; then artificially synthesize them with our team The standard viral nucleic acid and a large number of HPV-infected clinical biological samples were used as templates, and a large number of repeated polymerase chain reactions were performed using the above-mentioned pairs of candidate primers, and the obtained products were sequenced to verify the specificity and sensitivity of the amplification; finally, the specificity 30 pairs of primers with excellent sensitivity, the nucleotide sequences of the primers are as shown in SEQ ID No: 1 to SEQ ID No: 60.

[0025] The specific screening method includes the following steps.

[0026] (1) Randomly select a group of candidate primer pairs and carry out amplification reaction in a centr...

Embodiment 2

[0029] A method for detecting nucleic acid sequences of candidate primers for detecting HPV1 types.

[0030] (1) Randomly select a group of candidate primer pairs for HPV1 types, and perform amplification reaction in a centrifuge tube. The reaction system is 25 μl. The specific reaction system is 0.2 μM each for the forward primer and reverse primer, SYBR Green I, and 0.8 μM for dNTP , pfu DNA polymerase 2.5 U, 10× pfu Buffer 5 μl, 1 ng / μl plasmid DNA; set up a set of negative controls at the same time, use ddH to ddH the plasmid DNA in the above reaction system 2 Replace with O, and the rest of the components are the same; put the centrifuge tube into the fluorescent PCR reaction, set the temperature at 94 °C, pre-denature for 5 min, and then enter the following cycle: 94 °C for 50 s, 49 °C for 50 s, 72 °C for 1 min, 40 cycles, 72 ℃ for 5 min; then pick 3 sets of candidate primers and repeat the amplification reaction, from the obtained amplification curve, the primer sc...

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Abstract

The invention belongs to the field of microbiological detection, and particularly relates to a nucleic acid sequence for typing detection of cutaneous human papillomavirus and application thereof. The 30 pairs of primers can be used for preparing a diagnostic product for rapidly detecting a cutaneous HPV type, for example, a kit. The nucleotide sequence can rapidly and accurately detect the type of the cutaneous HPV and can meet the clinical requirements for typing detection of the cutaneous HPV.

Description

technical field [0001] The invention belongs to the field of detection of microbial pathogens, and in particular relates to a nucleic acid sequence for typing and detection of skin type human papillomavirus and its application. Background technique [0002] Human papillomavirus (Human papillomavirus, HPV) is a kind of papillomavacuovirus A belonging to Papovaviridae family. It is a spherical DNA virus that can cause squamous epithelial proliferation of human skin and mucous membranes. Symptoms include common warts and genital warts (condyloma acuminata). With the rapid increase in the incidence of genital warts among sexually transmitted diseases and the increase in cervical cancer and anal cancer, human papillomavirus (HPV) infection has attracted more and more attention. [0003] HPV widely exists in the natural environment and is closely related to human health. People began to explore more and more HPV-related diseases. At present, more than 200 kinds of HPV have been ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/708C12Q1/6844C12Q2600/112C12Q1/68C12N15/11C12Q1/70
Inventor 高兴华杜昱光葛格毛瑞齐瑞群孙明陈洪铎
Owner THE FIRST HOSPITAL OF CHINA MEDICIAL UNIV
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