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Primer group and detection system for detecting HPV common hypotypes through LAMP method

A primer set and system technology, used in recombinant DNA technology, microbial assay/inspection, biochemical equipment and methods, etc., to achieve high amplification efficiency, excellent specificity, and improved amplification efficiency.

Inactive Publication Date: 2017-07-11
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, there are no reports on the primer set and detection system for detecting common subtypes of HPV in the present invention.

Method used

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  • Primer group and detection system for detecting HPV common hypotypes through LAMP method
  • Primer group and detection system for detecting HPV common hypotypes through LAMP method
  • Primer group and detection system for detecting HPV common hypotypes through LAMP method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1 Materials and methods

[0037] 1.1 Experimental materials

[0038] 1.1.1 Sample related information

[0039] From July 2014 to October 2016, 627 outpatients and inpatients in Shanghai Tenth People's Hospital and Shanghai Tenth People's Hospital Branch were selected as genital wart symptoms. There was no statistical difference in age and gender, and all kinds of other skin inflammatory infections were excluded. All patient samples were selected before surgery or drug treatment. The warts excised by the patient, the exfoliated dander obtained from the skin surface, or the swabs of the genital tract were eluted and stored in 3ml of HPV sample preservation solution.

[0040] 1.1.2 Collection of samples for HPV typing

[0041] The above 624 samples were all tested for HPV infection and typing in the samples by using the perspective flow fluorescence hybridization method. Single samples of HPV-6, HPV-11, HPV-16, HPV-42, HPV-43, HPV-44 were selected. Samples with highe...

Embodiment 2

[0207] The target gene sequence used is as follows HPV-6 (HG793938.1); HPV-11 (KU298879.1); HPV-16 (KP313775.1); HPV-42 (KU298897.1); HPV-43 (HE962401.1); HPV-44 (HE963128.1). Design four primer sets of F3, B3, FIP, BIP.

[0208] A total of 164 cases of single positive samples were screened by perspective flow fluorescence hybridization method. Including 30 cases of HPV-6 type, 30 cases of HPV-11 positive, 22 cases of HPV-16 positive, 20 cases of HPV-42 positive, 30 cases of HPV-43 positive, 20 cases of HPV-44 positive. According to the operating instructions of the Loopamp DNA amplification kit, the HPV typing of different primer sets were detected respectively. The results are judged by the positive rising peak of the Loopamp real-time turbidimeter and the color change of HNB to blue, and the positive results are all in line with the positive characteristics. At the same time, a negative control was added, and the negative control was normal as the prerequisite for the ex...

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Abstract

The invention relates to a primer group and detection system for detecting HPV common hypotypes through a LAMP method. The HPV common hypotypes include HPV6, HPV11, HPV16, HPV42, HPV43 and HPV44. The primer group is composed of a primer sequence having a nucleotide sequence of SEQ NO.1-24. The primer group is designed according to the six hypotypes of condyloma acuminate and HPV infected by common skin warts, and the LAMP primers are verified. Through the reaction system above, the specific HPV hypotype DNA in a template can be effectively amplified. Through verification of specificity and sensitivity of the system, compared with a method for clinical application in the prior art, the LAMP system has superior sensitivity. Therefore, the invention provides a HPV type detection method that is simple and fast and is convenient in detection and high in specificity and sensitivity.

Description

technical field [0001] The invention relates to the technical field of molecular detection, in particular to a primer set and a detection body for detecting common subtypes of HPV by a LAMP method. Background technique [0002] Human papillomavirus (HPV) is a double-stranded circular DNA virus that can specifically infect the skin and mucous membranes of human reproductive organs, causing condyloma acuminatum (CA) and squamous intraepithelial lesions (SILs). Squamous intraepithelial lesions can further lead to cervical cancer. At present, more than 100 subtypes of HPV virus have been found, which can be divided into two types according to whether they can cause cervical cancer. Those that are closely related to cervical cancer and other external genital cancers and can cause high-grade cervical intraepithelial lesions are high risk types, mainly including HPV16, 18, 33, 35, 39, 45, etc. It is mainly related to genital warts, and low-risk types (Lowrisk Types) that cause ot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/708C12Q2600/112C12Q2531/119
Inventor 刘维薇钟琦陈丹刘妍李可
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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