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HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit

A detection kit and the technology of the kit, applied in the field of human papillomavirus high-risk type typing fluorescent PCR detection kit

Active Publication Date: 2014-09-03
SANSURE BIOTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The purpose of the present invention is to solve the defects of existing high-risk human papillomavirus nucleic acid typing detection kits, and provide a high-risk human papillomavirus fluorescent quantitative PCR assay with fast operation, simple method, high detection sensitivity and wide detection range. High-risk HPV types 16, 18, 31, 33, Type 35, Type 39, Type 45, Type 51, Type 52, Type 53, Type 56, Type 58, Type 59, Type 66, Type 68. The DNA nucleic acid fragments of these 15 types are subjected to rapid fluorescent PCR for typing detection. The test results can be used for auxiliary diagnosis of high-risk human papillomavirus infection, early screening of cervical cancer, follow-up of cervical lesions, and guidance for vaccine development

Method used

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  • HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit
  • HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit
  • HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit

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Experimental program
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Effect test

Embodiment 1

[0083] This embodiment provides a detection kit capable of detecting 15 high-risk human papillomaviruses and accurately typing them, which at least consists of the following components:

[0084] ① Nucleic acid release agent: Surfactin 0.01-0.5mmol / L (mass / volume), potassium chloride (KCl) 50-200mmol / L (mass / volume), sodium dodecylsulfonate (SDS) 0.01%~2% (mass / volume), ethanol 0.05%~1% (volume / volume);

[0085]②PCR reaction solution: the first PCR reaction solution containing HPV16 and 18 primer probe sequences, used to detect HPV16 and 18 types; the second PCR reaction solution containing HPV39 and 31 primer probe sequences, used For the detection of HPV39 and 31 types; the third PCR reaction solution containing HPV33 and 58 primer probe sequences, used to detect HPV33 and 58 types; the fourth PCR reaction solution containing HPV45 and 59 primer probe sequences Reaction solution for detecting HPV45 and 59 types; the fifth PCR reaction solution comprising HPV51 and 66 primer ...

Embodiment 2

[0094] The operation steps of using the high-risk human papillomavirus nucleic acid typing detection kit in Example 1 to detect high-risk HPV-DNA in unknown samples such as wart surface exfoliated cells, women's cervical epithelial cells, and reproductive tract secretions are:

[0095] 1. Reagent preparation

[0096] Take out each component in the kit, place it at room temperature, wait for its temperature to balance to room temperature, mix well and set aside;

[0097] Take n parts of 8 kinds of HPV PCR reaction solutions, each part is 38-44 μl, and 8×n parts of enzyme mixture, each part is 1-2 μl, mix well to make PCR-mix, centrifuge briefly and set aside. Wherein, n=number of samples to be tested+2 quality control substances, and the quality control substances are negative control and positive control. In addition, since the PCR reaction solution containing HPV16 and 18 type primer probes needs to be used as a quantitative reference product, n+4 copies of this PCR reaction...

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Abstract

The invention provides an HPV (human papilloma virus) high-risk typing fluorescence PCR (polymerase chain reaction) detection kit which comprises a nucleic acid releaser and two or more PCR reaction solutions, wherein the nucleic acid releaser comprises 0.01-0.5 mmol / L surfactin, 20-300 mmol / L potassium chloride, 0.01-2% of sodium dodecylsulfate and 0.05-1% of ethanol. One of the PCR reaction solutions comprises high-risk HPV 16 type and / or 18 type primer probe sequences, and another PCR reaction solution comprises an internal reference primer probe sequence. The kit provided by can be used for carrying out quick fluorescence PCR typing detection on DNA (deoxyribonucleic acid) nucleic acid segments of high-risk HPV in wart surface shedding cells, female cervix epithelial cells, genital secretion and other unknown samples; and the detection result can be used for auxiliary diagnosis of high-risk HPV infection, early screening of cervical carcinoma, follow-up survey of cervix pathological changes and instruction of vaccine development.

Description

technical field [0001] The invention provides a high-risk type typing fluorescent PCR detection kit for human papillomavirus (HPV). Background technique [0002] Human papillomavirus (HPV) is a kind of non-enveloped double-stranded circular DNA virus with small molecular weight, which obligately infects and parasitizes the epithelial cells of human reproductive organs and other tissues and organs. Clinically, HPV can be divided into high-risk and low-risk types according to the different pathogenicity or cancer risk of different HPV subtypes. Low-risk HPV mainly causes exophytic wart-like lesions on the anus skin, male external genitalia, female labia, urethral meatus, lower vagina, and low-grade cervical intraepithelial neoplasia. High-risk HPV can not only cause external genital warts, but more importantly, external genital cancer, cervical cancer and high-grade cervical intraepithelial neoplasia. The virus subtypes mainly include HPV16, 18, 31, 33, 35, 39, and 45 , 51, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/686C12Q1/70C12Q2523/113C12Q2531/113C12Q2545/101
Inventor 戴立忠刘佳李勃邓中平
Owner SANSURE BIOTECH INC
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