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Primer pair for detecting novel coronavirus SARS-CoV-2, probe, composition and kit and application thereof

A coronavirus and detection reagent technology, applied in the field of primer pairs for the detection of the new coronavirus SARS-CoV-2, can solve the problems of expensive instruments, difficulty in moving forward the epidemic prevention and control threshold, and inability of patients to cause pathogens, so as to meet the needs of rapid diagnosis and treatment. The effect of monitoring the whole process, controlling the epidemic, gaining time, and shortening the testing time

Pending Publication Date: 2020-06-12
齐永
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AI Technical Summary

Problems solved by technology

[0003] The real-time fluorescent RT-PCR (rRT-PCR) method currently relied on for diagnosis requires expensive instruments and professional operation, and it is difficult to promote and apply at the grassroots level
Front-line doctors in Wuhan once reported that "the biggest shortcoming is the virus nucleic acid detection kit", "the use conditions are strict, many people will not use it, it takes a long time, and the process is complicated, which is the bottleneck"; CCTV feedback "patients cannot carry out timely pathogen detection. leading to cross-infection”
This also makes it difficult to move forward the epidemic prevention and control gate, and a large backlog of suspected cases increases the risk of nosocomial infection and cross-infection

Method used

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  • Primer pair for detecting novel coronavirus SARS-CoV-2, probe, composition and kit and application thereof
  • Primer pair for detecting novel coronavirus SARS-CoV-2, probe, composition and kit and application thereof
  • Primer pair for detecting novel coronavirus SARS-CoV-2, probe, composition and kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1, be used to detect the design of the fluorescent RT-RAA primer of novel coronavirus SARS-CoV-2, probe

[0050] For the specific conserved region of the new coronavirus SARS-CoV-2 as the target region, specific primers and fluorescent RAA probes were designed. The sequences are:

[0051] Forward primer, such as the nucleotide sequence shown in SEQ ID No.: 1 in the sequence listing:

[0052] 5'-CCAGTGCTCAAAGGAGTCAAATTACATTAC-3'

[0053] Reverse primer, such as the nucleotide sequence shown in SEQ ID No.: 2 in the sequence listing:

[0054] 5'- AAAACAGCAAGAAGTGCAACGCCAACAATA -3'

[0055] Oligonucleotide probes, such as the nucleotide sequence shown in SEQ ID No.: 3 in the sequence listing:

[0056] 5'-GGTGAAATCAAGGATGCTACTCCTTCAGA[BHQ-dT][THF][FAM-dT]TGTTCGCGCTACTG-3';

[0057] Wherein: FAM-dT represents a thymidine deoxynucleotide carrying a fluorescein group FAM (6-Carboxyfluorescein);

[0058] THF represents a tetrahydrofuran (tetrahydrofuran) linker; ...

Embodiment 2

[0060] Embodiment 2. Establishment of a fluorescent RT-RAA method for detecting novel coronavirus SARS-CoV-2

[0061] (1) Fluorescence RT-RAA reaction

[0062] 1) Extraction of sample RNA: The sample RNA can be extracted by using the viral RNA nucleic acid extraction kit from Kaiser Bioengineering Co., Ltd., according to the kit instructions.

[0063] 2) Using the forward primer, reverse primer and probe designed in Example 1, and the RT-RAA reaction kit (this example specifically uses the RT-RAA basic kit of Jiangsu Qitian Gene Biotechnology Co., Ltd.), Using the sample RNA to be detected prepared in step 1) of this embodiment as a template, the amplification reaction is carried out, wherein the reaction system is as follows:

[0064] 25 μl RT-RAA reaction buffer (provided by RT-RAA basic kit of Jiangsu Qitian Gene Biotechnology Co., Ltd.);

[0065] 2.1 μL each of the forward primer and reverse primer (10 μM) designed in Example 1;

[0066] 0.6 μL of the probe designed in ...

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Abstract

The invention provides a primer pair for detecting novel coronavirus SARS-CoV-2, a probe, a composition, a kit and an application thereof. The fluorescent RT-RAA primer and the fluorescent probe can be used for rapidly and qualitatively detecting the novel coronavirus SARS-CoV-2 in a patient sample. According to the fluorescent RT-RAA primer and the fluorescent probe provided by the invention, when the fluorescent RT-RAA primer and the fluorescent probe are used for detecting the novel coronavirus SARS-CoV-2, the detection time is remarkably shortened compared with a PCR method or fluorescentPCR, and the sensitivity is equivalent to or even higher than that of the fluorescent PCR.

Description

technical field [0001] The invention relates to the technical field of in vitro nucleic acid detection, in particular to a primer pair, probe, composition, kit and application for detecting the novel coronavirus SARS-CoV-2. Background technique [0002] Since December 2019, the outbreak of infectious disease (COVID-19) caused by the novel coronavirus SARS-CoV-2 has posed a huge threat to social stability and people's health. In the face of the complex situation of the epidemic situation, the prevention and control gates have been moved forward to community medical centers and township health centers to solve the problems of infection and cross-infection of nosocomial infections on the way to medical treatment for patients. "Early detection" is an important means to deal with the epidemic. Faced with solving the technical problems of nucleic acid detection in grassroots units. [0003] The current real-time fluorescent RT-PCR (rRT-PCR) method that diagnosis relies on needs e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/701C12Q1/6844C12Q2521/507C12Q2522/101C12Q2563/107Y02A50/30
Inventor 齐永李越希谭伟龙吕瑞辰陆年宏艾乐乐沈万鹏朱长强韩一芳李文豪李银吕恒
Owner 齐永
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