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Universal virus sample preserving fluid and preparation method thereof

A preservation solution and general-purpose technology, applied in the field of biomedicine, can solve the problems of lack of virus sample preservation solution, etc., to reduce risks, avoid degradation, and ensure accuracy

Active Publication Date: 2022-02-08
NANJING VAZYME MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is a lack of a general-purpose inactivated virus sample preservation solution, which can meet the sample requirements for nucleic acid detection and colloidal gold detection at the same time

Method used

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  • Universal virus sample preserving fluid and preparation method thereof
  • Universal virus sample preserving fluid and preparation method thereof
  • Universal virus sample preserving fluid and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: the preparation of virus preservation solution

[0042] The first virus preservation solution (referred to as "preservation solution 1"): includes the following components: 100mmol / L of trishydroxymethylaminomethane hydrochloride, 12mmol / L of tetrasodium iminodisuccinate, and 1w / L of sucrose. v%, 0.9w / v% of sodium chloride, 0.2w / v% of sodium lauryl sulfate, 0.8w / v% of Pluracare 1307, 20mmol / L of ethylenediaminetetraacetic acid, 0.8 of casein % w / v, the liquid biological preservative Proclin 300 is 0.2w / v%, the solvent is RNase-free purified water, and the pH of the preservation solution is 8.0.

[0043] The second virus preservation solution (referred to as "preservation solution 2"): includes the following components: 100mmol / L of trishydroxymethylaminomethane hydrochloride, 5mmol / L of tetrasodium iminodisuccinate, and 2w / L of sucrose. v%, sodium chloride 1.5w / v%, sodium dodecyl sulfate 0.2w / v%, Pluracare 1307 1w / v%, ethylenediaminetetraacetic acid 30mmo...

Embodiment 2

[0046] Example 2: Nucleic acid detection and verification of nucleic acid preservation effect in virus preservation solution

[0047] 1) Add the inactivated virus culture of the new coronavirus (2019-nCOV) to the three kinds of sample preservation solutions in Example 1, and dilute it to the following concentration gradient: 800 TCID 50 、400TCID 50 、200TCID 50 、100TCID 50 、50TCID 50 、25TCID 50 . The Ct value corresponding to each concentration was obtained by amplifying with an approved and listed nucleic acid detection kit (PCR fluorescence method).

[0048] 2) Extraction of viral RNA: Take 200 μL of each sample with the above concentration gradient, and perform nucleic acid extraction according to the instructions of the viral nucleic acid extraction kit (Nanjing Novizan Biotechnology Co., Ltd., Cat.RC311-C1). The final viral RNA purification product elution volume was 50 μL.

[0049] 3) RT-qPCR fluorescence quantitative detection: configure the PCR reaction system ac...

Embodiment 3

[0060] Example 3: Colloidal gold detection and verification of virus preservation solution virus protein preservation effect

[0061] 1) The virus culture with a concentration gradient prepared by diluting the same batch of virus preservation solution in Example 2 above was tested using the protocol of the colloidal gold detection of the new crown antigen (Nanjing Novizan Medical Technology Co., Ltd., Cat.C8602CA). Observe the test card 10 minutes after the start of the test, and judge the result. After 15 minutes, the observation result is invalid.

[0062] 2) The test results shall be judged as follows:

[0063] Negative result: only a red quality control line (C line) can be seen by naked eye.

[0064] Positive result: Two clear red lines can be seen by naked eyes, one is the quality control line (C line) and the other is the T test line.

[0065] Invalid result: no red line visible to the naked eye or only T test line but no quality control line (C line), indicating that...

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Abstract

The invention belongs to the technical field of biomedicine, and relates to a general inactivated virus sample preserving fluid related to a virus detection reagent and a preparation method thereof. According to the virus sample preserving fluid, in the virus cracking process, the virus functional protein structure is not damaged, degradation of viral nucleic acid by endogenous nuclease can be avoided, and a sample preserved by the preserving fluid can be used for viral nucleic acid detection and also can be used for virus antigen or antibody detection.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a universal inactivated virus sample preservation solution related to virus detection reagents and a preparation method thereof. Background technique [0002] A new type of virus strain was first discovered in the human body in 2019. The novel coronavirus (Severe Acute Respiratory Syndrome Coronavirus 2, or SARS-CoV-2) is still raging around the world since its discovery, and is currently the most mainstream and most contagious virus. Respiratory virus. Since the virus is highly contagious during the incubation period, the infection rate caused by secondary contact remains high. For a large group of potential patients and close contacts, early detection, early reporting, early isolation, and early treatment are the basic prevention and control measures. move. Therefore, whether carrying the new coronavirus antigen has become a key clinical indicator for judging whether the te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806G01N33/53
CPCC12Q1/6806G01N33/5306C12Q2527/125C12Q2527/127Y02A50/30
Inventor 曹林唐波杨博文贺海涛
Owner NANJING VAZYME MEDICAL TECH CO LTD
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