A kit for detecting hpv16/18 based on digital PCR for accurate quantitative typing and its detection method

A quantitative typing and kit technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problem of no typing kit for HPV16/18 detection, and achieve result interpretation Intuitive and reliable, good accuracy and reproducibility, simplified operation steps

Active Publication Date: 2018-05-15
ZYBIO INC
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  • Abstract
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  • Claims
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Problems solved by technology

However, there is currently no kit and detection method for the accurate quantitative typing of HPV16 / 18 based on digital PCR

Method used

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  • A kit for detecting hpv16/18 based on digital PCR for accurate quantitative typing and its detection method
  • A kit for detecting hpv16/18 based on digital PCR for accurate quantitative typing and its detection method
  • A kit for detecting hpv16/18 based on digital PCR for accurate quantitative typing and its detection method

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Embodiment Construction

[0040] In order to better understand the content of the present invention, the present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the following specific examples are only used to illustrate the present invention, rather than limit the present invention.

[0041] 1. A kit for detecting HPV16 / 18 based on digital PCR accurate quantitative typing, including DNA extraction solution, digital PCR reaction buffer A, digital PCR reaction buffer B, HPV16 virus gene positive quality control, HPV18 virus gene positive quality control, negative quality control and internal standard solution;

[0042] The DNA extract comprises 50 mM Tris-HCl with pH 8.3, 0.5 M NaCl, 5 mM EDTA, CTAB with a mass concentration of 1.5%, and DTT with 2 mM;

[0043] The digital PCR reaction buffer A includes primers and probes of the HPV16 type and the primers and probes of the HPV18 type; the digital PCR reaction...

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Abstract

The invention discloses an HPV (human papilloma virus) 16 / 18 detection kit based on digital PCR (polymerase chain reaction) accurate quantitative typing and a detection method thereof. The kit comprises a DNA (deoxyribonucleic acid) extracting solution, a digital PCR reaction buffer solution A, a digital PCR reaction buffer solution B, an HPV 16 virus gene positive quality control, an HPV 18 virus gene positive quality control, a negative quality control and an internal standard solution. The method comprises the following steps: 1) treating a detected sample; 2) treating a quality control product; 3) preparing a digital PCR reaction mixed solution; 4) generating micro-reaction solution drops, and carrying out digital PCR reaction amplification; and 5) reading a fluorescent signal, analyzing the HPV type and calculating the number of copies. The method is independent of the external standard substance and standard curve, is simple to operate, and can directly perform accurate absolute quantitative simultaneous typing on HPV. The method has high sensitivity, and can reduce the interference of the background DNA and matrix and quantify trace HPV; and the detection limit is down to 1 copy.

Description

technical field [0001] The invention belongs to the technical field of molecular diagnostic biology, and in particular relates to a kit for detecting HPV16 / 18 based on digital PCR for accurate quantitative typing and a detection method thereof. Background technique [0002] Human papillomavirus (human papillomavirus HPV) is very common in human infection, can invade human epithelial cells, cause benign and malignant tumors of skin and mucous membrane tissue. More and more evidence shows that the consequences of human infection with HPV are closely related to the type of infected HPV, and different subtypes have specific infection sites and lesions, of which more than 40 subtypes are related to human reproductive tract diseases. According to its risk, it can be divided into high-risk type and low-risk type. Low-risk HPV types such as HPV-6, 11, 40, 42, 43, 44, 54, 61, 67, etc. mainly cause genital warts and flat warts of the genital tract, perianal skin, and extravaginal les...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/6851C12R1/93
CPCC12Q1/6851C12Q1/708C12Q2531/113C12Q2545/101C12Q2545/113C12Q2563/159
Inventor 廖璞吉权陈环
Owner ZYBIO INC
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