COVID-19 nucleic acid releasing agent and nucleic acid detection kit

A technology of COVID-19 and nucleic acid release agent, which is applied in the direction of DNA preparation, recombinant DNA technology, microbial measurement/inspection, etc. It can solve the problems of short detection time, inability to achieve rapid diagnosis, long detection process, etc.

Pending Publication Date: 2020-10-23
GUANGZHOU JINQIRUI BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these kits have good detection sensitivity and accuracy, the entire detection process is long and cannot achieve the purpose of rapid ...

Method used

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  • COVID-19 nucleic acid releasing agent and nucleic acid detection kit
  • COVID-19 nucleic acid releasing agent and nucleic acid detection kit
  • COVID-19 nucleic acid releasing agent and nucleic acid detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] A COVID-19 nucleic acid release agent, comprising: a surfactant with a mass volume percent concentration of 0.02-0.05%, and a buffer reagent with a pH value of 7.0-8.0;

[0066] The surfactant is selected from: NP40, X-100 and Tween20.

[0067] Include Tris-HCl, KCl, NaCl, DTT, (NH 4 ) 2 SO 4 and spermidine.

[0068] The concentration of the KCl is 40mM~50mM, the concentration of the NaCl is 5~8mM; the concentration of the DTT is 1~1.5mM; the (NH 4 ) 2 SO 4 The concentration of the spermidine is 5-8mM; the concentration of the spermidine is 1-1.5%.

Embodiment 2

[0070] A COVID-19 nucleic acid detection kit, comprising:

[0071] 1) the nucleic acid releasing agent of embodiment 1;

[0072] 2) Specific primer pairs and specific fluorescent probes for amplifying the virus sequence ORF1 region, N protein sequence, and monitoring the endogenous reference gene RnaseP of the detection process, the working concentration of the above primer pairs and probes is 10 μM ( 10 μmol / L), as shown in the table below.

[0073] Table 1. Specific primer pairs and specific fluorescent probe primer sequences

[0074] Primer / Probe Name Sequence (5'-3') serial number ORF1a-F AATGGTCATGTGTGGCGGTTCAC SEQ NO.1 ORF1a-R CAGCTTGACAAATGTTAAAAACACTA SEQ NO.2 ORFN-F CAGGCAGCAGTAGGGGAACTTCT SEQ NO.4 ORFN-R GGCCTTTACCAGACATTTTGCTCTCA SEQ NO.5 RNaseP-F CTGTGTGTCCACTCAGGCTTGT SEQ NO.7 RNaseP-R ACAGATGGGTCTCAGGTGCAG SEQ NO.8 ORF1a-P CTCATCAGGAGATGC CACAACTGCT SEQ NO.3 ORFN-P CTGCTCTTGCTTTG...

Embodiment 3

[0078] Adopt the kit of embodiment 2, carry out nucleic acid detection to COVID-19, method flow is as follows:

[0079] 1) Viral nucleic acid release:

[0080] Take 10 μL of a throat swab sample, add 10 μL of nucleic acid release agent, mix well, and let stand at room temperature for 10 minutes.

[0081] 2) RT-PCR system preparation:

[0082] Add the sample (20 μL) processed in step 1) to the RT-PCR reaction system, the RT-PCR reaction is 50 μL, including 5 μL of the primer / probe combination (each primer and probe in Table 1 of Example 3), 2 μL of enzyme solution (reverse transcriptase and Tag enzyme), 23 μL of reaction buffer (remaining components in the RT-PCR reaction reagent in Example 2); the RT-PCR reaction system was dispensed into 96-well plates.

[0083] 3) Put the 96-well plate containing the RT-PCR reaction system prepared in step 2) into the Roche 480 or ABI7500 instrument, set the reaction program, and start real-time detection.

[0084] 4) Result analysis: jud...

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Abstract

The invention relates to a COVID-19 nucleic acid releasing agent and a nucleic acid detection kit, and belongs to the technical field of gene detection. The COVID-19 nucleic acid releasing agent comprises a surfactant with the mass volume percentage concentration of 0.01-1% and a buffer reagent with the pH value of 6.0-8.8. According to the nucleic acid releasing agent, through cooperation of thesurfactant and the buffer reagent, on the basis that RNA of COVID-19 is not damaged, potential COVID-19 viruses of a sample to be detected can be split, nucleic acid is released into a solution system, and nucleic acid extraction can be replaced with two hours. Therefore, the whole detection process is shortened to be within 1.5 hours from four hours, and the COVID-19 nucleic acid releasing agenthas the advantages of being rapid in detection, high in sensitivity, high in specificity and good in reliability.

Description

technical field [0001] The invention relates to gene detection technology, in particular to a COVID-19 nucleic acid release agent and nucleic acid detection kit. Background technique [0002] COVID-19 is an acute infectious pneumonia whose pathogen is a novel coronavirus (COVID-19) not previously found in humans. The virus has strong infectivity and pathogenicity, and has spread globally. The infection rate and death rate are increasing, which seriously threatens people's lives and economic losses due to the impact of the virus spread on various industries. Therefore, diagnosis and detection of 2019-nCoV can help curb the development of the epidemic. [0003] Coronaviruses are enveloped RNA viruses that are widely found in human, mammalian, and avian hosts and can cause respiratory, intestinal, hepatic, and neurological diseases. For the detection of the new coronavirus, there are now immunology-based antibody detection kits and next-generation sequencing detection reagent...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12N15/10
CPCC12Q1/701C12Q1/686C12N15/1003C12Q2521/107C12Q2563/107C12Q2561/101
Inventor 朱方何任胜强邹树勇陈嘉昌柳俊胡朝晖
Owner GUANGZHOU JINQIRUI BIOTECHNOLOGY CO LTD
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