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Virus preserving fluid, kit and ultrasensitive detection method of virus RNA

A storage solution and kit technology, applied in the biological field, can solve the problems of large storage solution volume, reduced detection sensitivity, excessive sample dilution, etc., and achieve the effect of improving extraction efficiency.

Pending Publication Date: 2020-11-03
深圳柏悦基因科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Traditionally, the storage solution used to preserve virus samples is large in volume, and it is easy to over-dilute the sample, resulting in a decrease in the content of viral RNA, thereby reducing the sensitivity of detection

Method used

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  • Virus preserving fluid, kit and ultrasensitive detection method of virus RNA
  • Virus preserving fluid, kit and ultrasensitive detection method of virus RNA
  • Virus preserving fluid, kit and ultrasensitive detection method of virus RNA

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] This embodiment provides a virus preservation solution, which is composed of: 6M guanidine salt, 1% N-lauroyl sarcosine and 0.6M sodium citrate.

Embodiment 2

[0048] This embodiment provides a kit for virus RNA extraction, which contains several extraction units, each of which is composed of: 500ul of the virus preservation solution provided in Example 1, 2mg of nano-magnetic beads, and 300ul of binding buffer , washing buffer I 200ul, washing buffer II 200ul, 80% ethanol 200ul, eluent 50ul; wherein:

[0049] The composition of the binding buffer is: absolute ethanol;

[0050] The composition of washing buffer I is: 1.2M guanidinium salt, 60% ethanol;

[0051] The composition of washing buffer II is: 70% ethanol;

[0052] The composition of the eluent is: ultrapure water.

Embodiment 3

[0054] This embodiment provides a method for extracting viral RNA.

[0055] 1. The virus sample is obtained by the following method: wipe the oral mucosa with a clean sterile cotton swab, wipe it repeatedly 4 times, then spray the fake virus solution evenly on the cotton swab with a watering can, and put the cotton swab back into the preservation tube containing the preservation solution During storage, tighten the bottle cap and store at -20°C to 4°C for later use.

[0056] 2. The method for extracting viral RNA is as follows:

[0057] (1) Take 500ul of the collected sample in an RBX centrifuge tube (no exogenous DNA, RNA contamination), centrifuge at 10000rpm for 1min, and take all the filtrate;

[0058] (2) Add 300 ul of binding buffer to the filtrate obtained in step (1), shake and mix for 1 min, then add 2 mg of nano-magnetic beads and pipette to mix;

[0059] (3) Heat at 56°C for 10 minutes, mix once every 2-3 minutes during this period, place the centrifuge tube in a ...

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Abstract

The invention relates to virus preserving fluid, a kit for virus RNA extraction and an ultrasensitive detection method of virus RNA. According to the invention, a guanidine salt-containing solution isused for preserving a virus sample, so that the virus RNA can be well preserved for a long time, and the extraction efficiency of the virus RNA in the sample is improved. The detection method provided by the invention can realize ultrasensitive detection of virus RNA, and is of great significance to large-scale screening of viruses in crowds.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a virus preservation solution, a kit and an ultrasensitive detection method for virus RNA. Background technique [0002] Viral RNA is easily degraded and cannot survive for a long time. Since the virus cannot be detected in time after sampling, the virus RNA needs to be processed, and it needs to be stored in a virus preservation solution. Otherwise, the viral nucleic acid will degrade quickly and the nucleic acid cannot be detected, so it is necessary to add a preservation solution to the sampling tube to prevent the viral nucleic acid from being degraded so quickly. And ensure that the virus can be transported and retained. [0003] Due to the outbreak of the novel coronavirus in 2020, a large number of people need to undergo nucleic acid testing. If it takes a lot of time and energy to test samples one by one, it does not meet the actual requirements. Therefore, it is often ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/70C12R1/93
CPCC12Q1/6806C12Q1/701C12Q2527/125C12Q2523/308C12Q2523/32
Inventor 聂棱黄小浩
Owner 深圳柏悦基因科技有限公司
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