Alcohol-free virus nucleic acid extraction kit based on magnetic bead method

A virus nucleic acid and kit technology, applied in the field of bioengineering, can solve the problems of complex nucleic acid extraction methods, difficult purification, and high extraction requirements, and achieve the effects of significant technological progress, broad application prospects, and simple experimental steps.

Pending Publication Date: 2019-05-07
SHANGHAI KEHUA BIO ENG
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the above-mentioned technical problems in the prior art, the invention provides a kind of alcohol-free virus nucleic acid extraction kit based on the magnetic bead method, and the described alcohol-free virus nucleic acid

Method used

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  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method
  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method
  • Alcohol-free virus nucleic acid extraction kit based on magnetic bead method

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0027] Example 1 Detection of HBV and HCV viruses in plasma (compared with a magnetic bead method kit from a domestic manufacturer)

[0028] (1) Extraction of viral nucleic acid

[0029] 1) Add 320ul of plasma to a 2ml centrifuge tube (in negative plasma, add WHO HBV standard with a final concentration of 10IU and WHO HCV standard with a final concentration of 30IU), add 600μL of lysate, 15μL of solution containing magnetic beads, and 20μL of protease K solution and 5μL polyA solution, vortex for 1 min until the sample is well mixed, then shake and mix for 15 min.

[0030] 2) After instant centrifugation, place the centrifuge tube on the magnetic stand and let it stand for 1-2 min. After the magnetic beads are completely adsorbed, the liquid is aspirated.

[0031] 3) Add 500 μL of buffer solution to the protein-removing solution and mix well, vortex for 1 min.

[0032] 4) After instant centrifugation, place the centrifuge tube on the magnetic stand and let it stand for 1-2 min. After t...

Example Embodiment

[0053] Example 2 Detection of HBV and HCV virus in serum (compared with Shanghai Kehua Nucleic Acid Extraction Kit (lot: 18076550), operation steps refer to the instructions)

[0054] (1) Extraction of viral nucleic acid

[0055] 1) Add 320ul of plasma to a 2ml centrifuge tube (in negative plasma, add WHO HBV standard with a final concentration of 10IU and WHO HCV standard with a final concentration of 30IU), add 600μL of lysate, 15μL of solution containing magnetic beads, and 20μL of protease K solution and 5μL polyA solution, vortex for 1 min until the sample is well mixed, then shake and mix for 15 min.

[0056] 2) After instant centrifugation, place the centrifuge tube on the magnetic stand and let it stand for 1-2 min. After the magnetic beads are completely adsorbed, the liquid is aspirated.

[0057] 3) Add 500 μL of buffer solution to the protein-removing solution and mix well, vortex for 1 min.

[0058] 4) After instant centrifugation, place the centrifuge tube on the magnetic ...

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Abstract

The present invention provides an alcohol-free virus nucleic acid extraction kit based on a magnetic bead method. The alcohol-free virus nucleic acid extraction kit comprises a lysate, a magnetic beadsolution, proteinase K, a polyA solution, a deproteinized solution, a washing solution and a buffer solution. The present invention also provides a method for extracting nucleic acid by using the above kit. Firstly, a sample, the lysate and magnetic beads are mixed evenly, then nucleic acid molecules are captured while pyrolysis is conducted, and finally the special alcohol-free deproteinized solution and the washing solution are respectively used to remove impurities of proteins, salt ions and other impurities on surfaces of the magnetic beads. Compared with a conventional virus nucleic acidextraction method, the method is simple in operation steps, does not need single addition of binding liquid, realizes the simultaneous pyrolysis and binding, and co-extracts DNA and RNA; the whole operation process is free of alcohol or isopropyl alcohol, inhibitory substance residues are less, the experimental procedure is simple, and the whole process only takes about 40 minutes; and the method is safe and pollution-free, and free of toxic reagents of chloroform, phenol, etc., and can be operated manually or on an automation platform.

Description

technical field [0001] The invention belongs to the field of bioengineering and relates to a detection kit, in particular to an alcohol-free viral nucleic acid extraction kit based on a magnetic bead method. Background technique [0002] The structure of the virus is relatively simple, consisting of the outer capsid protein and the inner nucleic acid substance. According to the nucleic acid substance, the virus can be divided into DNA virus or RNA virus. With the development of molecular biology, the detection and typing of disease-associated viruses is an important application field of molecular detection, and the efficient and simple extraction and purification of target nucleic acids from biological materials is the key technology of molecular detection. Currently, viral nucleic acid purification technologies are mainly divided into three types, namely traditional method, column membrane method and magnetic bead method. [0003] Traditional nucleic acid purification main...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12N1/06
Inventor 王金辉方琴张健蒋淼
Owner SHANGHAI KEHUA BIO ENG
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