LAMP (loop-mediated isothermal amplification) primer group and kit for bedside rapid detection of HPV (human papillomavirus) and use method of LAMP primer group

Abstract

The invention discloses a bedside LAMP (loop-mediated isothermal amplification) primer group for rapidly detecting HPV (human papilloma virus), a kit and a use method of the kit, and belongs to the technical field of biological detection. Firstly, an LAMP primer group for detection is provided, so that the detection specificity, sensitivity and the like can be effectively ensured; secondly, the primer is applied to preparation of an HPV detection kit or reagent, DNA extraction and amplification are combined into a one-step method through optimization of the optimal proportion of system solution components, LAMP reaction time is shortened, the total reaction time is shortened to be within 40 min, and meanwhile high sensitivity and excellent specificity are shown; finally, aiming at the kit, the invention provides a use method of the bedside rapid HPV detection kit, and the use method specifically comprises the steps of sample collection, splitting decomposition, amplification, discrimination and the like. In addition, in consideration of the problem of environmental pollution, the inventor also specially designs a closed waste liquid collecting cavity to achieve the purpose of eliminating pollution.

Description

technical field [0001] The invention relates to a primer set, a kit and a use method for detecting HPV, in particular to a LAMP primer set, a kit and a use method for bedside rapid detection of HPV, and belongs to the technical field of biological detection. Background technique [0002] Human papillomavirus (HPV) is a papilloma vacuolar DNA virus belonging to the Papovaviridae family. It can spread widely in the human body. Continuous infection can lead to skin and mucous membrane vegetations, genital warts and cervical cancer. Therefore, Therefore, rapid and accurate detection of HPV is of great significance to reduce the incidence of cervical cancer. Traditional HPV detection methods include: cytopathological detection, colposcopy detection, electron microscopy detection and cervical fluoroscopy, etc. These detection methods have obvious disadvantages: low sensitivity, poor specificity, and inability to type HPV, resulting in the inability to be used in clinical practice....

AI Technical Summary

Problems solved by technology

Traditional HPV detection methods include: cytopathological detection, colposcopy detection, electron microscopy detection and cervical fluoroscopy, etc. These detection methods have obvious disadvantages: low sensitivity, poor specificity, and inability to type HPV, resulting in the inability to be used in clinical practice. on regular use

At present, the routine clinical HPV detection methods are mainly: real-time fluorescent PCR, hybridization capture-chemiluminescence and PCR capillary electrophoresis fragment analysis, etc., but there are disadvantages such as requiring professional equipment and professional operation, and taking a long time.

[0003] The prior art "CN110607399A, primer combinations, kits and methods for detection of HPV and typing by LAMP amplification" discloses HPV16E6, HPV18E6, HPV31E6, HPV33E7, HPV35E6, HPV39E6, HPV45E7, HPV51E6E7, HPV52E6E7, HPV56E6, HPV58E6 , HPV59E6, HPV66E6, HPV68E6, and HPV73E6E7 and other LAMP amplification primer sets, using the above primer combinations to detect HPV subtypes, but this method has the following disadvantages: (1) DNA needs to be extracted, which takes more than 2 hours; (2) LAMP amplification Increase reaction for 1 hour

[0004] In addition, only HPV16 and HPV18 are detected in "CN104805218A Reaction System and Method for Detecting HPV16 and 18 Based on LAMP and Molecular Beacon", and this method has the following disadvantages: (1) The reaction system contains molecular beacon probes, which are relatively expensive High; (2) It takes a long time, more than 60 minutes; (3) After the LAMP reaction, it needs to be irradiated with a UV lamp to observe the fluorescence at 4°C, 65°C and 95°C. The operation is cumbersome and requires professional equipment: UV lamp; " CN106148571A detects human papillomaviruses HPV16 and HPV18 complete set of reagents "in detection time is longer, about 1 hour, and needs ultraviolet light; " CN106939359A a kind of LAMP method detects the primer group and detection system of HPV common subtype " detects HPV6 , HPV11, HPV16, HPV42, HPV43 and HPV44, the detection method involved takes a long time, more than 1 hour; special equipment is required, such as: fluorescent quantitative PCR instrument, vortex mixer, oscillating constant temperature metal bath, high-speed low-temperature centrifuge , electrophoresis instrument, gel imaging analyzer, flow matrix analyzer, real-time turbidimeter, electronic analytical balance, ultrapure water instrument, etc.; "CN110093459A LAMP primer combination and application for rapid detection of 13 high-risk HPV" It takes more than 1 hour, and during the reaction, it is necessary to use a turbidimeter to detect turbidity and draw an "S" curve

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