Nucleic acid releasing agent and HPV virus nucleic acid detection kit

Abstract

The invention discloses a nucleic acid releasing agent. The nucleic acid releasing agent comprises a composition of Triton X-100 and diethylene glycol monoethyl ether (DCDE) with a mass ratio of 1: (1-3). The low-cost composition of Triton X-100 and DCDE is adopted to replace a traditional guanidine salt and a detergent, and the obtained nucleic acid releasing agent has advantages of safe use, easy degradation, simple operation, and low manufacturing cost, can effectively lyse serum samples containing pathogens, quickly release nucleic acids therein, reduce inhibition of downstream experiments, and has high pathogen lysis efficiency. The invention also discloses an HPV virus nucleic acid detection kit. The kit comprises the nucleic acid releasing agent, is simple and convenient to operate,and high in working efficiency, effectively avoids a link that may occur during the experimental operation, reduces loss of HPV DNA in an extraction process, and has a quantification limit of 45 IU / mL and a detection limit of 22 IU / mL.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid release agent and an HPV virus nucleic acid detection kit. Background technique [0002] Real-time fluorescent quantitative PCR detection of free virus in blood is the most advanced clinical detection method at this stage, and it has developed quite maturely, but the extraction steps of viral nucleic acid before detection are very cumbersome, such as: (1) Traditional boiling method: alkali The neutral lysate is directly mixed with the pathogen-containing liquid, lysed at high temperature, and centrifuged to obtain nucleic acid; (2) concentrated boiling method: first use polysaccharide to concentrate and precipitate pathogens, then add alkaline lysate to lyse at high temperature, and centrifuge to obtain nucleic acid; (3) centrifuge Column method: use silica gel membrane or glass fiber membrane to absorb, purify, and elute to obtain nucleic acid; (4) Magnetic bead metho...

AI Technical Summary

Problems solved by technology

For example, Chinese patent 201310009055.4 discloses a one-step pathogen nucleic acid fluorescent quantitative PCR detection method, selects a conserved gene sequence for the pathogen nucleic acid to be tested, designs a specific primer probe, uses a nucleic acid rapid release reagent to extract the pathogen nucleic acid, and directly adds the corresponding The PCR reaction solution realizes PCR fluorescence quantitative detection. The invention is simple and fast in operation, has less loss of extracted nucleic acid, and the manufacturing cost of reagents used is low, but its nucleic acid rapid release agent still has an impact on the subsequent PCR amplification detection, and cannot affect serum low Accurate quantification of HPV DNA up to 400 copies / mL

Images