Hepatitis B virus nucleic acid extraction reagent and extraction method thereof

Abstract

The invention discloses a hepatitis B virus nucleic acid extraction reagent and an extraction method thereof. The extraction reagent is prepared from the following raw materials in parts by weight: 20to 40 parts of reagent A, 15 to 30 parts of reagent B, 15 to 30 parts of reagent C, 50 to 80 parts of reagent D, and magnetic nanoparticles; the reagent A is prepared from the following materials: Tris , EDTA, NP-40, sodium chloride, 3-hydroxypropane sulfonic acid, fatty alcohol polyoxyethylene ether sodium sulfate, guanidinium isothiocyanate, beta-mercaptoethanol, and water; the reagent B is prepared from the following raw materials: guanidine hydrochloride, water saturated phenol, ammonium sulfate, xylitol, pyrrolidone, isobutanol; the reagent C s prepared from the following raw materials:dimethylformamide and tartronic acid; the reagent D is anhydrous ethanol; and the magnetic nanoparticles are nanometer iron oxide. The hepatitis B virus nucleic acid extraction reagent and the extraction method thereof can extract low concentration free nucleic acid at room temperature without using proteinase K and carrier RNA, and has the advantages that operation is simple, extraction efficiency and extraction purity are improved.

Description

technical field [0001] The invention relates to the field of gene technology, in particular to a hepatitis B virus nucleic acid extraction reagent and an extraction method thereof. Background technique [0002] In the quantitative detection of clinical hepatitis B virus (HepatitisBVirus, HBV) nucleic acid, HBVDNA extraction is the first step of operation. At present, the nucleic acid extraction methods mainly include phenol-chloroform extraction, spin column method and magnetic bead method. Phenol-chloroform extraction is highly toxic to the operator due to the use of a large amount of toxic solvents, and it is difficult to realize automatic operation. Due to the low efficiency and high cost of free nucleic acid extraction, the spin column method requires a high-speed centrifuge, and it is difficult to automate, so it cannot be widely used in large-scale clinical testing. The magnetic bead extraction nucleic acid technology developed in recent years uses superparamagnetic ...

AI Technical Summary

Problems solved by technology

However, the commercial reagents based on the magnetic bead method currently on the market are too complicated to operate, and need to be incubated with proteinase K and carrierRNA at a certain temperature, and multi-step washing steps are required, resulting in low extraction efficiency.