Method of extracting low-abundance viral nucleic acid

Abstract

A method of extracting low-abundance viral nucleic acid is disclosed. The method includes steps of: adding a lysis liquid into blood to release viral nucleic acid into the solution; adding a biotin-labeled specific nucleic acid fragment capable of hybridization with the viral nucleic acid; adding streptavidin-labeled nanometer magnetic beads, wherein streptavidin on surfaces of the magnetic beads can be combined with biotin to form a magnetic bead-streptavidin-biotin-specific nucleic acid fragment-viral nucleic acid composite; and washing the magnetic beads with a washing liquid to remove the viral nucleic acid from the magnetic beads. By adoption of the method, the extraction efficiency and the yield of nucleic acid in low-titer viral samples can be increased so as to reduce false negative detection results due to the low extraction efficiency. The method is particularly suitable for clinic blood screening, and other applications.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the use of nano magnetic beads to extract low-abundance viral nucleic acids from blood, including single-stranded and double-stranded DNA and RNA. Background technique [0002] Nucleic acid extraction and purification technology is a basic technology of biochemistry and molecular biology. As molecular biology technology is widely used in biology, medicine and related fields, nucleic acid extraction and purification technology has also been further developed. [0003] Nucleic acids are combined with various proteins in cells. The extraction of nucleic acid mainly refers to the separation of nucleic acid from protein, polysaccharide, fat and other biological macromolecular substances. Most nucleic acid extraction steps generally include cell lysis, removal of nucleic acid-bound proteins, polysaccharides, lipids and other biological components, removal of other unwanted nucleic ac...

AI Technical Summary

Problems solved by technology

[0007] Since the surface of ordinary nano magnetic beads is coated with a layer of silicate material, all nucleic acids can be adsorbed during the nucleic acid extraction and purification process, and a mixture of various nucleic acids is obtained after elution, which will affect the subsequent PCR amplification and other Interference caused by sample processing may also reduce the yield and yield of some low-abundance nucleic acid sequences. For clinical tests that require high sensitivity, it will cause false negative test results