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Nucleic acid extraction-free inactivated virus preserving fluid and application thereof

A technology of preservation solution and virus, which is applied in the direction of microbial measurement/inspection, biochemical equipment and methods, etc., and can solve the problems that viruses cannot be completely and effectively inactivated

Pending Publication Date: 2021-04-30
JIANGSU COWIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this preservation solution cannot completely effectively inactivate pathogenic microorganisms such as viruses, so samples need to be inactivated at high temperature (50-60°C) before nucleic acid detection to ensure the safety of experimental operators

Method used

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  • Nucleic acid extraction-free inactivated virus preserving fluid and application thereof
  • Nucleic acid extraction-free inactivated virus preserving fluid and application thereof
  • Nucleic acid extraction-free inactivated virus preserving fluid and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1. Virus sample preservation solution formula

[0059] The formula of the extraction-free virus sample preservation solution of the present invention mainly includes the following components: NaOH, proteinase K, Tris-HCl, Triton-100, EDTA, gelatin, Proclin-300 and β-propiolactone. Wherein the final concentration of Tris HCl is 10-200mM, preferably 50-100mM; the final concentration of Triton-100 is 0.01-1% (v / v), preferably 0.01-0.5%; the final concentration of EDTA is 0.5-5mM, preferably 0.5- 1mM; the final concentration of gelatin is 1-5%, preferably 2-4%; the final concentration of NaOH is 10-200mM, preferably 50-100mM; the final concentration of proteinase K is 10-30mg / L, preferably 10-20mg / L The final concentration of Proclin-300 is 0.01-0.5%, preferably 0.01-0.05%; the final concentration of β-propiolactone is 0.001-0.05%, preferably 0.001-0.005%; the pH value of virus preservation solution is 7-9, preferably pH value for 7-8.

[0060] Table 1. Recipe ...

example 2

[0062] Example 2. Study on the Inactivation Efficacy of Inactivated Direct Expansion Preservation Solution to Chicken Infectious Bronchitis Virus

[0063] Experimental group preservation solution: No. 1, 2, 3, and 4 virus preservation solutions in Example 1

[0064] Control group storage solution: the inactivated virus storage solution invented by patent CN111334503A (including 5mg / ml nuclease inhibitor EDTA, 40mg / ml protein denaturant SDS, 50mg / ml stabilizer glycine, 100mg / ml bacteriostatic agent potassium sorbate , 80mM Tricine salt buffer.)

[0065] Experimental materials: 10-day-old SPF chicken embryos (purchased from Yangzhou), IBV seed virus for the experiment (stored in a -80°C refrigerator).

[0066] Experimental method: the experimental setup is as shown in Table 2. The chicken infectious bronchitis virus IBV strain seed virus prepared in equal parts was added to the virus preservation solution of the experimental group and the control group respectively, according t...

example 3

[0074] Example 3. The research on the preservation ability of RNA virus by virus preservation solution

[0075] In the present embodiment, using avian IBV virus (coronavirus, RNA virus, which is not infective to humans) as a model, it is detected that the inactivated direct expansion type virus preservation solution provided by the present invention has the effect on RNA virus sample nucleic acid under different storage temperature conditions. preservation effect.

[0076] Experimental materials: No. 1, 2, 3, and 4 virus preservation solutions in Example 1, the experimental virus species IBV seed virus (preserved in a -80°C refrigerator).

[0077] Experimental method: Add 20 μl of IBV virus stock solution to No. 1, 2, 3, and 4 virus preservation solutions respectively, mix them upside down, divide each sample into three equal parts, store them at 37°C, 4°C, and -80°C respectively, and store them at Take it out on the 7th day, and take 10 μl of each sample as a template (the a...

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Abstract

The invention provides a nucleic acid extraction-free inactivated virus preserving fluid and application thereof, and particularly, the virus sample preserving fluid comprises protease K, sodium hydroxide, beta-propiolactone, gelatin and other components, and virus nucleic acid can be effectively released and preserved at room temperature through the synergistic cooperation of the components. Nucleic acid degradation is effectively prevented in the normal-temperature transportation and storage process of the sample, and the method can be used for preparing clinical viral nucleic acid samples.

Description

technical field [0001] The invention belongs to the field of molecular diagnosis, in particular, the invention relates to an inactivated virus preservation solution free from nucleic acid extraction and application thereof. Background technique [0002] The virus-infected pneumonia (COVID-19), which broke out at the end of 2019, has posed a huge threat to the health of people around the world and caused major economic losses. Rapid and accurate pathogenic diagnosis is an important prerequisite for effective treatment, condition monitoring and control of disease spread. At present, the screening and diagnosis of patients by real-time fluorescent quantitative polymerase chain reaction (qPCR) is the main means of prevention and control of new coronary pneumonia. How to ensure the timeliness and accuracy of nucleic acid test results is of great significance to the current epidemic prevention and control. [0003] The nucleic acid of RNA virus is one of the most difficult biomo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/24
CPCC12Q1/24
Inventor 殷剑峰刘淑君王春香
Owner JIANGSU COWIN BIOTECH CO LTD
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