SARS-CoV-2 D614G mutation detection kit and SARS-CoV-2 D614G mutation detection method

A technology of 1. sars-cov-2d614g, 2. sars-cov-2d614g, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of high price, long time-consuming whole genome sequencing, Unfavorable for large-scale work and other issues, to achieve the effect of short detection cycle, simple and fast operation, and high sensitivity

Active Publication Date: 2021-03-30
XIAMEN UNIV +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Whole-genome sequencing is not only time-consuming and expensive, but also requires specialized equipment and prof

Method used

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  • SARS-CoV-2 D614G mutation detection kit and SARS-CoV-2 D614G mutation detection method
  • SARS-CoV-2 D614G mutation detection kit and SARS-CoV-2 D614G mutation detection method
  • SARS-CoV-2 D614G mutation detection kit and SARS-CoV-2 D614G mutation detection method

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Embodiment Construction

[0025] 1. The kit primer probe of the present invention

[0026] The present invention selects the conserved region covering the D614G mutation (23403A->G) on the S gene of the novel coronavirus to design primers and probes. Design a detection probe for D614 and G614 respectively, wherein the 5' end of the D614 detection probe is labeled with a ROX fluorophore, and the 3' end is labeled with a quencher group; the 5' end of the G614 detection probe is labeled with a FAM fluorophore, and the 3' end Label the quenching group. After extensive screening of primer-probe combinations, the final primer and probe sequences are shown in the table below:

[0027] Table 1. Primer and probe sequences of SARS-CoV-2 D614G mutation detection kit

[0028]

[0029] Note: "+" refers to LNA modification.

[0030] 2. Sample Processing

[0031] Nasopharyngeal swabs, oropharyngeal swabs, anterior and middle nasal swabs, nasopharyngeal wash / nasal aspirate, or bronchoalveolar lavage (BAL) sampl...

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Abstract

The invention discloses an SARS-CoV-2 D614G mutation detection kit and an SARS-CoV-2 D614G mutation detection method. The sequences of a primer and a probe for SARS-CoV-2 D614G mutation nucleic acid detection are shown as SEQ ID NO: 1-4. According to the SARS-CoV-2 D614G mutation detection kit, asymmetric PCR amplification is adopted, a multicolor probe melting curve analysis technology is combined, typing detection of SARS-CoV-2 S-D614 and S-G614 strains can be achieved, and the kit has the advantages of being easy to operate, short in detection period, high in sensitivity and the like.

Description

technical field [0001] The invention relates to a SARS-CoV-2 D614G mutation detection kit. Background technique [0002] The novel coronavirus (Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2) belongs to the genus Betacoronavirus along with Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) and Middle East Respiratory Syndrome Coronavirus (MERS-CoV) . SARS-CoV-2 can cause a respiratory infection in humans, causing a deadly lung inflammation known as novel coronavirus pneumonia (COVID-19). COVID-19 is a new acute respiratory infectious disease, which is highly contagious among the population and is mainly transmitted through respiratory droplets, respiratory secretions and direct contact. [0003] SARS-CoV-2 is a positive-strand RNA virus with 5 essential genes encoding 4 structural proteins: nucleoprotein (N), viral envelope (E), matrix protein (M) and spike protein (S), and RNA-dependent RNA polymerase (RdRp). The nucleoprotein (N) wraps the RNA genome...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/6858C12N15/11
CPCC12Q1/701C12Q1/6858C12Q2600/166C12Q2531/107C12Q2563/107C12Q2545/113C12Q2527/107Y02A50/30
Inventor 张永有宋娜杰
Owner XIAMEN UNIV
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