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Preserving fluid for biological sample nucleic acid detection and application thereof

A biological sample and preservation solution technology, applied in the field of molecular biology, can solve the problems of high price, poor effect of preservation of white blood cells, and inapplicability of rapid diagnosis and detection, so as to improve accuracy and reliability, reduce preservation temperature and preserve The effect of time requirements

Pending Publication Date: 2020-08-18
WUXI SHENRUI BIO PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Qiagen's preservation solution is very effective in preserving tissues, but it is not effective in preserving cells, especially white blood cells. The extraction of nucleic acids in preserved samples requires additional special purification reagents, which are expensive and not suitable for rapid diagnostic testing in clinical medicine.

Method used

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  • Preserving fluid for biological sample nucleic acid detection and application thereof
  • Preserving fluid for biological sample nucleic acid detection and application thereof
  • Preserving fluid for biological sample nucleic acid detection and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041]The preservation solution consists of nuclease inhibitors, surfactants, chelating agents and buffers. Specifically, the nuclease inhibitor is selected from guanidine hydrochloride, the surfactant is selected from cetyltrimethylammonium bromide (CTAB), the chelating agent is selected from ethylenediaminetetraacetic acid sodium salt, and the buffer is selected from Tris-HCl buffer. The final concentration of the nuclease inhibitor is 1.0M, the final concentration of the surfactant is 1.0W / V%, the final concentration of the chelating agent is 2mM, and the final concentration of the buffer is 50mM. In this embodiment, a buffer solution is added to the preservation solution to adjust the pH value of the preservation solution to 6.5.

Embodiment 2

[0043] The preservation solution consists of nuclease inhibitors, surfactants, chelating agents and buffers. Specifically, the nuclease inhibitor is selected from urea, the surfactant is selected from sodium dodecyl sulfate (SDS), the chelating agent is selected from ethylenediaminetetraacetic acid potassium salt, the buffer is selected from phosphate buffer, and nuclease inhibition The final concentration of the surfactant is 4.0M, the final concentration of the surfactant is 0.1W / V%, the final concentration of the chelating agent is 9.5mM, and the final concentration of the buffer is 500mM. In this embodiment, a buffer solution is added to the preservation solution to adjust the pH value of the preservation solution to 6.0.

Embodiment 3

[0045] The preservation solution consists of nuclease inhibitors, surfactants, chelating agents and buffers. Specifically, the nuclease inhibitor is selected from the combination of urea guanidine and lithium chloride in any ratio, and the surfactant is selected from dodecyldimethylbenzyl ammonium chloride (DDBAC) and sodium dodecyl sulfate (SDS) Combination in any ratio, the chelating agent is selected from ethylene glycol diethyl ether diamine tetraacetic acid sodium salt, and the buffer is selected from citrate buffer. The final concentration of the nuclease inhibitor is 2M, the final concentration of the surfactant is 0.1W / V%, the final concentration of the chelating agent is 4.5mM, and the final concentration of the buffer is 500mM. In this embodiment, a buffer solution is added to the preservation solution to adjust the pH value of the preservation solution to 5.0.

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Abstract

The invention provides a preserving fluid for biological sample nucleic acid detection and an application thereof. The preserving fluid is composed of a nuclease inhibitor, a surfactant, a chelating agent and a buffer solution. The preserving fluid is mixed with a solid sample or a liquid sample to denature proteins, inactivate pathogens, and inactivate DNA and RNA nuclease. The pathogens are inactivated, so the safety of a potential high-infectivity clinical sample is greatly improved, and good protection is formed for surrounding personnel and downstream operating personnel of the sample; the DNA and RNA nuclease is inactivated, so that the content and integrity of DNA and RNA in the sample are kept in a sampling state to the maximum extent, and the strict requirements on the aspects oftemperature, time and the like of clinical sample transportation and storage are reduced. The preserving fluid can be used for clinical high-infectivity nucleic acid detection of biological samples, and the biological samples comprise throat swabs, body fluids, lavage fluids, whole blood, whole blood cells, serum or plasma.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a preservation solution for nucleic acid detection of biological samples and its application. Background technique [0002] Nucleic acid can be divided into ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) according to its chemical composition. As one of the carriers of life genetic information, ribonucleic acid (RNA) exists in cell organisms, some viruses, and viroids, and participates in various life activities. Therefore, it has been widely studied in the field of molecular biology in scientific research and clinical testing High-quality, complete RNA can be used in various molecular biology experiments and tests, such as nuclease protection experiments, detection of proto-oncogene expression, drug monitoring, etc. The single-stranded structure of RNA itself is unstable and easily degraded by ribonuclease (RNase), which exists widely and is extremely stable. I...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2527/125C12Q2527/127
Inventor 朱桂春于垚垚李涛姚鲁帅盛青松
Owner WUXI SHENRUI BIO PHARMA
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