HPV (human papilloma virus) 16/18 detection kit based on digital PCR (polymerase chain reaction) accurate quantitative typing and detection method thereof

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 l...

AI Technical Summary

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

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