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Nucleic acid extraction-free virus preservation solution

A preservation solution and nucleic acid-free technology, applied in the field of molecular biology, can solve problems such as difficulty in preservation solution, achieve the effect of avoiding extraction and wide application range

Inactive Publication Date: 2020-06-23
TIANDZ INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In view of this, the present invention provides a virus preservation solution that can not only preserve the virus, but also be directly used for nucleic acid amplification without nucleic acid extraction, thereby solving the technical problem that the preservation solution in the prior art is difficult to be directly used for nucleic acid amplification

Method used

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  • Nucleic acid extraction-free virus preservation solution
  • Nucleic acid extraction-free virus preservation solution
  • Nucleic acid extraction-free virus preservation solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Rabbit Hemorrhagic Disease Virus (RHDV) preservation solution (RNA virus) free of nucleic acid extraction

[0045] In this example, taking rabbit viral hemorrhagic virus as an example, it is stored in the nucleic acid extraction-free preservation solution provided in this example, and it is tested whether the preservation solution containing rabbit viral hemorrhagic virus can be directly used for nucleic acid amplification detection.

[0046] The components of the nucleic acid extraction-free preservation solution in this example are as follows: component A includes: 20nM acetic acid-sodium acetate buffer, pH 2.0; Tween 20, with a volume percent concentration of 0.2%; pepsin 5mg / mL;

[0047] Part B contains: 0.5M Tris-HCl buffer, pH 7.5.

[0048] Rabbit Viral Hemorrhage Virus was stored in Fraction A and kept at ambient temperature until use.

[0049] Amplify directly with the mixture of combination A and component B of the present invention: mix component A...

Embodiment 2

[0056] Embodiment 2 Canine parainfluenza virus preservation solution (RNA virus) free of nucleic acid extraction

[0057] In this example, taking canine parainfluenza virus as an example, it is stored in the nucleic acid extraction-free preservation solution provided in this example, the viability of the virus in the preservation solution of the present invention is detected, and the canine parainfluenza virus is detected and preserved. Whether the preservation solution of influenza virus can be directly used for nucleic acid amplification detection.

[0058] The components of the nucleic acid extraction-free preservation solution in this example are as follows: Component A includes: 0.1M acetic acid-sodium acetate buffer, pH 4.5; Tween 20, with a volume percent concentration of 2%; pepsin 100 mg / mL;

[0059] Part B contains: 1.0M Tris-HCl buffer, pH 8.9.

[0060] Canine parainfluenza virus was stored in Fraction A and kept at ambient temperature until use.

[0061] Direct a...

Embodiment 3

[0068] Example 3 African swine fever virus preservation solution (DNA virus) free of nucleic acid extraction

[0069] In this example, taking African swine fever virus as an example, it is stored in the nucleic acid extraction-free preservation solution provided in this example, the viability of the virus in the preservation solution of the present invention is detected, and the African pigs are detected Whether the pestivirus preservation solution can be directly used for nucleic acid amplification detection.

[0070] The components of the nucleic acid extraction-free preservation solution in this example are as follows: Component A includes: 50nM acetic acid-sodium acetate buffer, pH 3.5; Tween 20, with a volume percent concentration of 1%; trypsin 50mg / mL;

[0071] Part B contains: 0.7M Tris-acetic acid buffer, pH 8.

[0072] African swine fever virus was stored in Fraction A and kept at ambient temperature until use.

[0073] African swine fever virus nucleic acid amplif...

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Abstract

The present invention discloses a nucleic acid extraction-free virus preservation solution. The nucleic acid extraction-free virus preservation solution comprises a component A and a component B. Thecomponent A comprises an acidic buffer, Tween 20 and acidic protease; wherein, the molar concentration of the acidic buffer is 20 mM-100 mM, pH is 1.0-5.0, volume percentage of the Tween 20 is 0.1%-2%and concentration of the acid protease is 0.1-100 mg / mL; and the component B comprises a neutral or alkaline buffer, the concentration is 0.5-1 M and pH is 7.0-9.0. The preservation solution can be used to preserve RNA viruses and DNA viruses. During a preservation process, virus infectivity is severely weakened or even completely lost, and the preservation solution can be directly used for virusnucleic acid amplification without needing nucleic acid extraction.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a preservation solution that can not only preserve virus samples but also be directly used for nucleic acid amplification. Background technique [0002] Virus is a kind of non-cellular organism with relatively primitive life form and life characteristics, usually composed of genome and coat protein. Usually viruses have the following characteristics: smiling in shape and lack of cellular structure; containing only one nucleic acid, DNA or RNA; relying on its own nucleic acid for replication; lack of complete enzymes and energy systems; strict intracellular parasitism. From the classification of genetic material, viruses can be divided into DNA viruses, RNA viruses, and protein viruses (such as prions). From the perspective of virus structure, viruses can be divided into Euvirus (Euvirus, virus for short) and Subvirus (Subvirus, including viroid, pseudovirus, prion). I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12Q1/6844
CPCC12N7/00C12Q1/6844
Inventor 徐堤
Owner TIANDZ INC
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