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Reagent kit for detecting 14 high-risk HPV subtypes, and detection method

A detection method and kit technology, applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems of low accuracy, complicated operation, and large demand for detection samples, and achieve Guaranteed specific effect

Active Publication Date: 2020-07-28
天津普瑞赛斯分子诊断技术有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The sequencing method has low sensitivity (only 20%), which leads to high false negatives, and there are many steps and complicated operations
Fluorescent quantitative PCR method is currently the mainstream detection method, with high sensitivity and simple operation, but there are also shortcomings in the large demand for detection samples, low detection throughput and low accuracy.

Method used

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  • Reagent kit for detecting 14 high-risk HPV subtypes, and detection method
  • Reagent kit for detecting 14 high-risk HPV subtypes, and detection method
  • Reagent kit for detecting 14 high-risk HPV subtypes, and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] In this example, the plasmid constructed by genetic engineering was used as the positive plasmid, and the wild-type human genomic DNA was used as the control. 14 HPV subtypes and 70 HPV clinical samples were detected by using Highly Multiplexed PCR (Highly Multiplexed PCR) and high-precision capillary electrophoresis technology of the present invention, and compared with fluorescent quantitative PCR detection. The specific steps and methods are as follows:

[0085] 1. Test sample processing and DNA extraction:

[0086] (1) Plasmid treatment and extraction: each plasmid was extracted using a plasmid extraction kit, and the specific extraction steps were operated according to the kit instructions. The extracted plasmid was dissolved in Tris-EDTA (10mmol / L, pH 8.0), the quality of the extraction was detected by Nanodrop, and its concentration was determined, and then the concentration of the plasmid was adjusted to different copy numbers with Tris-EDTA (10mmol / L, pH 8.0),...

Embodiment 2

[0099]In this example, the plasmid constructed by genetic engineering was used as the positive plasmid, and the wild-type human genomic DNA was used as the control. 14 types of HPV and 70 cases of HPV clinical samples were detected by using Highly Multiplexed PCR (Highly Multiplexed PCR) and high-precision capillary electrophoresis technology of the present invention. The specific steps and methods are as follows:

[0100] 1. Test sample processing and DNA extraction:

[0101] (1) Plasmid treatment and extraction: each plasmid was extracted using a plasmid extraction kit, and the specific extraction steps were operated according to the kit instructions. The extracted plasmid was dissolved in Tris-EDTA (10mmol / L, pH 8.0), the quality of the extraction was detected by Nanodrop, and its concentration was determined, and then the concentration of the plasmid was adjusted to different copy numbers with Tris-EDTA (10mmol / L, pH 8.0), and used as PCR Template, take 5 μL for PCR reac...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a reagent kit for detecting 14 high-risk HPV subtypes, and a detection method. The reagent kit for detecting 14 high-risk HPV subtypes comprises HPV reaction liquid, HPV positive control and HPV negative control, wherein the HPV reaction liquid consists of primers and PCR buffer liquid, the HPV positive control isplasmid DNA, and the HPV negative control is wild type human genome DNA. The detection method is high in detection speed, the whole detection process including an extraction step only needs 150min, besides, the detection method has the characteristics of being good in specificity, low in price, high in accuracy and simple to operate, and actual requirements for clinical fast detection can be completely met.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a kit and a detection method for detecting 14 high-risk HPV types. Background technique [0002] HPV virus is the abbreviation of human papillomavirus, which is a papillomavacuolating virus A genus of Papovaviridae, and is a sexually transmitted disease caused by spherical DNA virus infection. Cervical cancer is second only to breast cancer in prevalence among women. There are about 130,000 new cases of cervical cancer and about 30,000 deaths each year in my country. Almost all cervical cancers are caused by high-risk HPV infection. Among women infected with high-grade lesions in China, the infection rate ranks among the top ten The most common HPV types are: 16, 52, 58, 18, 33, 31, 51, 53, 68, 59. It is very necessary to perform genotyping in HPV detection. Different HPV types cause different clinical manifestations, and can be divided into skin low-risk type, skin hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/708C12Q1/686C12Q2600/112C12Q2537/143C12Q2565/125
Inventor 王晨光王海瑶纪东华孙翼辰党晓萌
Owner 天津普瑞赛斯分子诊断技术有限责任公司
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