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Primer group and kit for detecting novel coronavirus SARS-CoV-2

A coronavirus, sars-cov-2 technology, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of complex CrRNA design, expensive RPA reagents, not suitable for widespread promotion, etc. To achieve the effect of simple equipment requirements, low cost and short time consumption

Active Publication Date: 2020-08-07
上海国际旅行卫生保健中心
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Problems solved by technology

[0006] For the new coronavirus, there are literature reports on the detection idea of ​​Cas13 enzyme combined with RPA technology, but because Cas13 enzyme has RNA cleavage activity, the operation is complicated, the required CrRNA design is also longer and more complicated, and the RPA reagent is expensive, which is not suitable for the market. Widely promoted

Method used

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  • Primer group and kit for detecting novel coronavirus SARS-CoV-2
  • Primer group and kit for detecting novel coronavirus SARS-CoV-2
  • Primer group and kit for detecting novel coronavirus SARS-CoV-2

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Embodiment 1

[0049] 1. Determination of the target gene sequence for detection of novel coronavirus

[0050] Compared the coronavirus strains from Wuhan, bat-SL-CoVZC45, Rs4231 of Chinese chrysanthemum bats, four other SARS virus sequences with the closest homology, human SARS epidemic strains in 2003, civet isolated sequences, common 4 human coronavirus sequences (229E, HKU1, NL63, OC43), etc., the results are as follows figure 1 As shown, the fragment and S gene of the new coronavirus orf1a / b were selected as the target gene for detection.

[0051] The nucleotide sequence of the novel coronavirus orf1a / b fragment is shown in SEQ ID No.15, specifically as follows:

[0052] AAGTAAATGAGTTCGCCTGTGTTGTGGCAGATGCTGTCAT AAAAACTTTGCAACCAGTATCTGAATTACTTACACCACTGGGC ATTGATTTAGATGAGTGGAGTATGGCTACATACTACTTATTTGA TGAGTCTGGTGAGTTTAAATTGGCTTCACATATGTATTGTTCTT TCTACCCTCCAGATGAGGATGAAGAAGAAGGTGATTGTGAAG AAGAAGAGTTTGAGCCATCAACTCAATATGAGTATGGTACTGA AGATGATTACCAAGGTAAACCTTTGGAATTTGGTGCCACTTCT GCTGCTCTTCAAC...

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Abstract

The invention provides a novel coronavirus SARS-CoV-2 detection primer group and a kit, and belongs to the technical field of virus detection. The primer group comprises a primer group aiming at an orf1a / b fragment and / or a primer group aiming at an S gene; the primer group aiming at the orf1a / b fragment comprises F3-inside, B3-inside, FIP-inside, BIP-inside, LF-inside and crRNA-inside-R, and theprimer group aiming at the orf1a / b fragment comprises F3-inside, B3-inside, FIP-inside, BIP-inside, LF-inside and crRNA-inside-R; and the primer group for the S gene comprises S gene-F3, S gene-B3, Sgene-FIP, S gene-BIP, S gene-LF, S gene-LB and crRNA-Sgene-R. The primer group and the kit are used for detecting the novel coronavirus, the consumed time is short, the cost is low, the detection sensitivity is high, and the specificity is good.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a primer set and a kit for detection of novel coronavirus SARS-CoV-2. Background technique [0002] In response to the epidemic, the country responded quickly and launched a series of prevention and control measures. And share the genetic sequence information of the virus with the international community after the discovery of the pathogen. Subsequently, the Chinese Center for Disease Control and Prevention announced the primers and probes for SARS-CoV-2 fluorescent PCR detection, and the fluorescent PCR method became the method for confirming the infection in this outbreak. However, in recent days, more and more on-site detection reactions, fluorescent PCR methods have detected false negatives. In addition, due to the complex operation of the fluorescent PCR method, professional personnel are required for detection, and the reaction time is long, which make...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2521/327Y02A50/30
Inventor 张子龙田桢干张威李深伟张晓航杜鹃张宏陆晔
Owner 上海国际旅行卫生保健中心
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