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Primer composition and detection method for detecting novel coronaviruses

A technology of primer composition and coronavirus, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., can solve the problems of high price and inability to widely develop nucleic acid detection technology, and achieve low cost, high efficiency, and specificity strong effect

Pending Publication Date: 2020-09-04
吴涛
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no matter whether it is to detect SARS-CoV-2 nucleic acid through a real-time fluorescent PCR platform or a gene sequencing platform, medical institutions or testing institutions are required to be equipped with expensive precision instruments (PCR analyzers and gene sequencers), complete laboratories Conditions and strictly trained professional and technical personnel
It is also limited by the above conditions that the current SARS-CoV-2 nucleic acid detection technology cannot be widely carried out in primary medical institutions, nor can it sink to most laboratories with biosafety level 2 laboratories and level 3 personal protection capabilities. Medical institutions and testing institutions

Method used

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  • Primer composition and detection method for detecting novel coronaviruses
  • Primer composition and detection method for detecting novel coronaviruses
  • Primer composition and detection method for detecting novel coronaviruses

Examples

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Embodiment 1

[0047] This embodiment provides a primer composition for detecting a novel coronavirus (SARS-CoV-2), including a primer set for detecting a novel coronavirus (SARS-CoV-2) by loop-mediated isothermal amplification technology, the primer set Including primer sets and / or nucleotides whose nucleotide sequences are SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 The sequence is the primer set of SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12.

Embodiment 2

[0049] This embodiment provides a kit for detecting coronavirus (SARS-CoV-2) using loop-mediated isothermal amplification technology, which contains the primer combination for detecting novel coronavirus (SARS-CoV-2) of Example 1 Things.

[0050] The kit uses loop-mediated isothermal amplification technology, and designs 8 pairs of specific primers with the conserved sequences in the ORF1ab gene and N gene of the novel coronavirus (SARS-CoV-2) as the target sites, among which F1c and F2 constitute the upstream The internal primers FIP (Forward Inner Primer), B1c and B2 constitute the downstream external primer BIP (Backward Outer Primer), the upstream loop primer LF (Loop Forward) and the downstream loop primer LB (Loop Backward) constitute a loop primer pair. The 3 primer pairs F3, B3, FIP, BIP and loop primer, under the action of Bst DNA polymerase with replication activity and high strand displacement activity, can achieve 6 regions of target gene sequence at a constant temper...

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Abstract

The invention discloses a primer composition and detection method for detecting novel coronaviruses, and particularly discloses a method for detecting novel coronaviruses by a loop-mediated isothermalamplification technique. An ORF1ab gene and an N gene in a complete genome of the novel coronaviruses are used as target genes, a specific primer is designed, and a reaction system is optimized, so that the target genes are amplified. The primer composition has the characteristics of being simple to operate, high in efficiency and low in cost. The primer composition has the characteristics of being high in specificity and convenient and swift when being used for detecting the novel coronaviruses, the detection efficiency can be improved, the detection requirements of a basal laboratory can bemet, and a new method is provided for detection of the novel coronaviruses.

Description

Technical field [0001] The invention relates to the technical field of virus detection, in particular to a primer composition and a detection method for detecting a novel coronavirus. Background technique [0002] At present, the positive nucleic acid of the new coronavirus (hereinafter referred to as SARS-CoV-2) is the most authoritative and direct pathogenic evidence for the diagnosis of COVID-19, and its detection mainly relies on real-time fluorescent RT-PCR technology and gene sequencing platforms. However, whether it is a real-time fluorescent PCR platform or a gene sequencing platform for SARS-CoV-2 nucleic acid detection, medical institutions or testing institutions need to be equipped with expensive precision instruments (PCR analyzers and gene sequencers) and complete laboratories Conditions and rigorously trained professional and technical personnel. It is also limited by the above-mentioned conditions that the current SARS-CoV-2 nucleic acid detection technology cann...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6844C12N15/11C12R1/93
CPCC12Q1/701C12Q1/6844C12Q2531/113C12Q2565/113C12Q2563/107
Inventor 吴涛
Owner 吴涛
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