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Kit for fluorescent PCR (Polymerase Chain Reaction) detection of high-risk HPV (Human Papilloma Virus)

A technology for detection kits and kits, which are used in the determination/inspection of microorganisms, microorganisms, biochemical equipment and methods, etc.

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 the existing high-risk human papillomavirus fluorescent quantitative PCR detection kit, and provide a high-risk human papillomavirus fluorescent quantitative PCR detection with fast operation, simple method, high detection sensitivity and wide detection range Reagent test kit

Method used

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  • Kit for fluorescent PCR (Polymerase Chain Reaction) detection of high-risk HPV (Human Papilloma Virus)
  • Kit for fluorescent PCR (Polymerase Chain Reaction) detection of high-risk HPV (Human Papilloma Virus)
  • Kit for fluorescent PCR (Polymerase Chain Reaction) detection of high-risk HPV (Human Papilloma Virus)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0083] This embodiment provides a detection kit capable of detecting 15 high-risk human papillomaviruses, 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: 5 μl of 10×PCR reaction buffer, 0.05mmol / L~0.2mmol / L deoxyribonucleoside triphosphate, 0.1μmol / L~0.3μmol / L upstream and downstream primers for target polynucleotide amplification HPV16-F and HPV16-R, HPV18-F and HPV18-R, HPV31-F and HPV31-R, HPV33-F and HPV33-R, HPV35-F and HPV35-R, HPV39-F and HPV39-R, HPV45- F and HPV45-R, HPV51-F and HPV51-R, HPV52-F and HPV52-R, HPV53-F and HPV53-R, HPV56-F and HPV56-R, HPV58-F and HPV58-R, HPV59-F and HPV59-R, HPV66-F and HPV66-R, HPV68-F and HPV68-R; 0.1 μmol / L~0.3 μmol / L probes for target polynucleotide detection HPV16-P, HPV18-P, ...

Embodiment 2

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

[0095] 1. Prepare reagents in the reagent preparation area

[0096] According to the number of samples to be tested, negative control, positive control, and quantitative reference products A~D, take the corresponding amount of PCR reaction solution (38~44μl / person) and enzyme mixture (1~2μl / person) in proportion and mix thoroughly. Homogenize PCR-mix, centrifuge briefly and set aside.

[0097] 2. Sample processing in the sample processing area

[0098] 1) Add 2-5 μl of nucleic acid release agent into each PCR reaction tube (it is recommended to inhale deeply and shallowly to avoid air bubbles), and add the sample to be tested, negative control, positive control, and quantitative reference product to different PCR reaction tubes. 3~5μl;

[0099] 2) At an interval...

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Abstract

The invention provides a kit for fluorescent PCR (Polymerase Chain Reaction) detection of a high-risk HPV (Human Papilloma Virus). The kit comprises a nucleic acid releasing agent and a PCR reaction solution, wherein the nucleic acid releasing agent comprises 0.01-05mmol / L surfactin, 20-300mmol / L potassium chloride, 0.01-2% sodium dodecyl sulfate and 0.05-1% ethanol; the PCR reaction solution comprises high-risk HPV16-type and 18-type primer probe sequences and internal standard primer probe sequences. The invention aims at providing a kit for fluorescent quantitative PCR detection of high-risk Human Papilloma Virus, and the kit has the advantages of fastness in operation, simple method, high detection sensitivity and wide detection range. By virtue of the kit, a common fast fluorescent PCR detection is carried out on DNA nucleic acid fragments of the high-risk human papilloma virus in unknown samples, such as cast-off cells of the surface of a verruca acuminata body, women cervical epithelial cells and genital secretion and the detection results can be applied in the auxiliary diagnosis of the high-risk human papilloma virus infection and the early screening of cervical cancer.

Description

technical field [0001] The invention provides a high-risk 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, 52, 53, 56, ...

Claims

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Application Information

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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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