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Virus sample treating fluid and treating method of novel coronavirus and rapid constant-temperature reverse transcription amplification kit for detecting virus

A sample processing liquid and coronavirus technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganism measurement/inspection, etc., can solve the problems of time-consuming, loss, and cost of nucleic acid extraction steps, and achieve nucleic acid detection Easy, small reaction volume, cost-saving effect

Pending Publication Date: 2021-01-08
杭州缔蓝生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the nucleic acid detection process, in addition to the amplification reaction, there is another important link is the extraction and purification of nucleic acid, especially for the extraction of RNA virus nucleic acid is very critical, because RNA is extremely unstable and easily degraded, after extraction It cannot be amplified directly, and it needs to be reverse transcribed into complementary DNA (cDNA) to be amplified. The nucleic acid extraction step is usually time-consuming, costly, and has a certain degree of loss. The efficiency of reverse transcription also varies with different methods. different
There is currently a lack of a quick and easy method for the detection of the novel coronavirus

Method used

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  • Virus sample treating fluid and treating method of novel coronavirus and rapid constant-temperature reverse transcription amplification kit for detecting virus
  • Virus sample treating fluid and treating method of novel coronavirus and rapid constant-temperature reverse transcription amplification kit for detecting virus
  • Virus sample treating fluid and treating method of novel coronavirus and rapid constant-temperature reverse transcription amplification kit for detecting virus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Virus inactivation method

[0082] Viruses are inactivated in two ways:

[0083] Method 1: Inactivate the virus by chemical method with a final concentration of 0.2M guanidine hydrochloride. The specific implementation process is as follows: take 960 μL of clinically collected virus sample solution, add 40 μL of 5.0 M guanidine hydrochloride solution, shake and mix for 10 seconds, and centrifuge at 13000 rpm for 1.5 minutes to obtain the processed sample to be tested.

[0084] Method 2 uses 0.1xTE buffer solution with a final mass percentage of Tween-20 of 0.01% to chemically inactivate the virus. The specific implementation process is as follows: take 960 μL of the clinically collected virus sample solution, add 40 μL of 0.1xTE buffer solution containing 0.25% Tween-20 by mass, treat it in a water bath at 95°C for 10 minutes, cool it, and centrifuge it at 13000rpm for 1.5min. The processed sample to be tested. The tube cap can only be opened after the inactivated vi...

Embodiment 2

[0086] Virus inactivation method

[0087] Viruses are inactivated in two ways:

[0088] Method 1: Inactivate the virus by chemical method with a final concentration of 0.4M guanidine hydrochloride. The specific implementation process is as follows: take 920 μL of clinically collected virus sample solution, add 80 μL of 5.0 M guanidine hydrochloride solution, shake and mix for 10 seconds, and centrifuge at 13,000 rpm for 1.5 minutes to obtain the processed sample to be tested.

[0089] Method 2 uses 0.1xTE buffer with a final mass percentage of Tween-20 of 0.1% to chemically inactivate the virus. The specific implementation process is as follows: take 600 μL of clinically collected virus sample solution, add 400 μL of 0.1xTE buffer solution containing 0.25% Tween-20 by mass, treat it in a water bath at 95°C for 10 minutes, cool it, and centrifuge it at 13000rpm for 1.5min. The processed sample to be tested. The tube cap can only be opened after the inactivated virus is coole...

Embodiment 3

[0091] Virus inactivation method

[0092] Viruses are inactivated in two ways:

[0093] Method 1: Use the chemical method to inactivate the virus with a final concentration of 0.6M guanidine hydrochloride. The specific implementation process is as follows: take 880 μL of clinically collected virus sample solution, add 120 μL of 5.0 M guanidine hydrochloride solution, shake and mix for 10 seconds, and centrifuge at 13000 rpm for 1.5 minutes to obtain the processed sample to be tested.

[0094] Method 2 uses 0.1xTE buffer solution with a final mass percentage of Tween-20 of 0.05% to chemically inactivate the virus. The specific implementation process is as follows: take 800 μL of clinically collected virus sample solution, add 200 μL of 0.1xTE buffer solution containing 0.25% Tween-20 by mass, treat it in a water bath at 95°C for 10 minutes, cool it, and centrifuge it at 13000rpm for 1.5min. The processed sample to be tested. The tube cap can only be opened after the inactiva...

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Abstract

The invention relates to a virus sample treating fluid and treating method of novel coronavirus and a rapid constant-temperature reverse transcription amplification kit for detecting the novel coronavirus, and belongs to the technical field of pathogenic microorganism in-vitro molecular detection. The virus sample treating fluid of the novel coronavirus is a guanidine hydrochloride aqueous solution with the final concentration of 0.1-1 M or a 0.1xTE buffer solution containing Tween-20 with the final mass percentage of 0.01%-0.1%. A virus sample treated with the virus treating fluid can realizesimple, convenient and rapid virus nucleic acid detection, and a result can be observed only by constant-temperature amplification and color development methods.

Description

technical field [0001] The present invention relates to the technical field of in vitro molecular detection of pathogenic microorganisms, in particular to a virus sample treatment solution and processing method of a novel coronavirus (2019-nCoV) and a rapid isothermal reverse transcription amplification (Quick Isothermal reverse transcription Amplification) for detecting the novel coronavirus , Q-IRTA) kit. Background technique [0002] The new coronavirus 2019-nCOV is an RNA virus that mainly infects mammals. Humans are one of its hosts. It first invades the upper respiratory tract, from the nasal cavity, nasopharynx, oropharynx, down to the hypopharynx, trachea, and bronchi. , alveolar cells, until the entire lung tissue is infected, causing lung inflammation. According to research, the main pathogenic mechanism of the new coronavirus is to cause a strong response from the host's immune system. A large number of inflammatory cells gather, release cytokines, and then cause...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6806C12Q1/6844C12R1/93
CPCC12Q1/701C12Q1/6806C12Q1/6844C12Q2527/125C12Q2521/107C12Q2521/101
Inventor 赵民清李萌方园任云
Owner 杭州缔蓝生物技术有限公司
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