Nucleic acid releasing agent and hepatitis B virus (HBV) nucleic acid releasing method

Abstract

The invention discloses a nucleic acid releasing agent and a hepatitis B virus nucleic acid releasing method. The nucleic acid releasing agent comprises 0.02 to 0.2 vol.% of Gemini quaternary ammoniumsalt surfactant, 50 to 300 mM of potassium hydroxide, isopropanol (0.2-2 mL / 100 mL), and the balance being sterile water. The nucleic acid releasing agent with a trace amount can rapidly destroy theprotein structure of the shell of pathogen, encloses and precipitates substances that can influence the PCR amplification such as serum proteins, and makes HBV DNA carry out complexing so as to completely precipitate out HBV DNA. The releasing agent is safe to use, the operation is simple and convenient, and the manufacturing cost is low. The releasing agent is used to release HBV DNA of a serum,blood plasma, or whole blood sample without heating or centrifugation; only through one-step sample loading, DNA can be released and extracted. According to the method, HBV nucleic acid extraction andamplification-fluorescence PCR quantitative detection can be carried out in one step, and the problems of loss and contamination of nucleic acid in a multi-step nucleic acid extraction method are solved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a nucleic acid release agent and a hepatitis B virus nucleic acid release method. Background technique [0002] At present, there are mainly four methods for nucleic acid extraction: (1) traditional boiling method: alkaline lysate is directly mixed with pathogen-containing liquid, pyrolyzed, and centrifuged to obtain nucleic acid. (2) Concentrated boiling method: first concentrate and precipitate pathogens with polysaccharides, then add alkaline lysate to lyse at high temperature, and centrifuge to obtain nucleic acids. (3) Spin column method: use silica gel membrane or glass fiber membrane to absorb, purify, and elute to obtain nucleic acid. (4) Magnetic bead method: use silica powder or polyethylene-diene-coated magnetic beads to absorb, purify, and elute to obtain nucleic acids. [0003] Method (1)(2) requires many steps such as changing tubes and centrifugation during extractio...

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Problems solved by technology

[0003] Method (1)(2) requires many steps such as changing tubes and centrifugation during extraction, the sample processing time is long, and the yield is low; the spin column method is better than method (1)(2), but the steps are still Various, low extraction efficiency; magnetic bead method has simple extraction steps, high recovery rate, and easy automatic extraction, but the product price is very expensive and is not widely used at present

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