Inactivated virus preservation solution and preparation method thereof

A technology for preserving fluids and viruses, applied in biochemical equipment and methods, and microbial measurement/inspection, etc., can solve the problems of increasing the difficulty of washing and removal, guanidine salt residue, secondary transmission of infection, etc.

Pending Publication Date: 2021-05-11
苏州赛普生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, in vitro diagnosis can be carried out by collecting samples such as throat swabs, nasal swabs, and blood from patients to detect whether they are infected with a certain virus. Common detection methods are antigen antibody detection or nucleic acid detection on patient samples, but the virus generally leaves the host. Cells will be inactivated quickly, the integrity of the virus will be destroyed, the virus shell will disintegrate, and the nucleic acid will be degraded. Therefore, in order to ensure the accuracy of in vitro diagnosis, the transport and preservation of virus samples require specific equipment or preservation solutions.
[0003] At present, the main virus preservation solutions on the market are generally physiological saline, PBS buffer or UTM transport medium. of biological samples are at risk of secondary transmission of infection
[0004] At present, most inactivated virus preservation solutions on sale are inactivated with high-concentration guanidine salts. Although high-concentration guanidine salts have a better effect on virus inactivation, high-concentration guanidine salts are prone to crystallization at low temperatures (for example, samples collected at transportation in low-temperature areas in the north in winter), the inactivation effect of the virus may be reduced after precipitation, and the risk of infection transmission during transportation is increased; Residues of guanidinium salt in nucleic acid, which will inhibit nucleic acid amplification, lead to missed detection and increase the proportion of false negative samples

Method used

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  • Inactivated virus preservation solution and preparation method thereof
  • Inactivated virus preservation solution and preparation method thereof
  • Inactivated virus preservation solution and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Preparation of inactivated virus preservation solution: Weigh 59.08 g of guanidine isothiocyanate, 15.76 g of trishydroxymethylaminomethane hydrochloride, 6.74 g of tetrasodium iminodisuccinate, 5 g of sodium dodecylsulfonate, benzene Add 0.5 g of ammonium chloride, 1 g of dithiothreitol, add 800 mL of purified water, adjust the pH value to 6.0 with NaOH, add purified water to make the volume to 1 L, so that the final concentration of guanidine isothiocyanate is 0.1 mol / L, three Hydroxymethylaminomethane hydrochloride is 100mmol / L, tetrasodium iminodisuccinate is 30mmol / L, sodium dodecylsulfonate is 0.50% (w / v) by weight and volume, benzalkonium chloride is The weight to volume ratio is 0.05% (w / v), and the weight to volume ratio of dithiothreitol is 0.10% (w / v). After preparation, the solution was sterilized through a 0.25 μm filter.

[0029] 1. Verification of virus inactivation performance

[0030] In the experimental group, the lentivirus with high-brightness gree...

Embodiment 2

[0032] Example 2 Virus Nucleic Acid Preservation Effect and Nucleic Acid Detection Verification

[0033] 1 Sample processing: Get each experimental batch by the virus preservation solution prepared in Example 1, marked as Cellpro 3 tube (2mL / tube) and the virus preservation solution prepared according to the prior art (wherein, the final concentration of sodium citrate The final concentration of proteinase K is 25mmol / L, the final concentration of proteinase K is 200μg / mL, the final concentration of concentrated sulfuric acid is 2.5% (V / V), the final concentration of ethylenediaminetetraacetic acid is 30mmol / L, and the final concentration of ammonium sulfate is 60%. (W / V)) marked as CompA 3 tubes (2mL / tube), add the RNA pseudovirus sample (Goldwise Biotechnology Co., Ltd.) of serial dilution, make the concentration successively 10 3 copies / mL, 10 4 copies / mL, 10 5 copies / mL, store the preservation solution added with pseudovirus samples at room temperature (20-25°C, humidity...

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PUM

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Abstract

The invention relates to an inactivated virus preservation solution and a preparation method thereof. The inactivated virus preservation solution comprises guanidinium isothiocyanate, trihydroxymethyl aminomethane hydrochloride, tetrasodium iminodisuccinate, an anionic surfactant, a cationic surfactant and dithiothreitol. According to the inactivated virus preservation solution provided by the invention, not only the integrity of viral nucleic acid can be effectively guaranteed , but also the components for inactivating viruses are added, so that the inactivation performance of the viruses is guaranteed, a collected biological sample can be applied to nucleic acid detection, the risk of virus transmission infection is reduced, and subsequent nucleic acid extraction and amplification detection are not influenced.

Description

technical field [0001] The invention relates to an inactivated virus preservation solution and a preparation method thereof, belonging to the technical field of in vitro diagnosis. Background technique [0002] Viruses are non-cellular life forms that must parasitize and proliferate in living cells. They have a simple structure and are generally composed of nucleic acid (DNA or RNA) and a protein shell that wraps the nucleic acid. Most viruses are pathogenic microorganisms that can cause disease in the host and can spread in the organism to be infectious. At present, in vitro diagnosis can be carried out by collecting samples such as throat swabs, nasal swabs, and blood from patients to detect whether they are infected with a certain virus. Common detection methods are antigen antibody detection or nucleic acid detection on patient samples, but the virus generally leaves the host. Cells will be inactivated quickly, the integrity of the virus will be destroyed, the virus she...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125
Inventor 张胜有叶竹青
Owner 苏州赛普生物科技股份有限公司
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