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Digital PCR (Polymerase Chain Reaction) detection kit for novel coronavirus and variants thereof

A coronavirus, variant strain technology, applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, DNA/RNA fragments, etc., can solve problems such as large demand for samples

Active Publication Date: 2021-07-06
领航医学科技(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, ARMS-PCR requires multiple tests to determine the corresponding subtype, and requires a large number of samples

Method used

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  • Digital PCR (Polymerase Chain Reaction) detection kit for novel coronavirus and variants thereof
  • Digital PCR (Polymerase Chain Reaction) detection kit for novel coronavirus and variants thereof
  • Digital PCR (Polymerase Chain Reaction) detection kit for novel coronavirus and variants thereof

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Embodiment 1

[0030] This embodiment discloses a method for simultaneously detecting multiple novel coronaviruses and variants thereof, including:

[0031] Based on the genome sequence of the novel coronavirus, primer express software was used to design primer probes for N gene, ORF1ab gene, S gene HV69-70del, K417N, K417T, N501Y mutants and human endogenous internal control genes.

[0032] The specific information is shown in Table 1 below.

[0033] Table 1

[0034]

[0035]

[0036]

[0037] 2. Samples

[0038] (1) ORF1ab gene and N gene pseudoviruses: purchased from biological companies, using RNA extraction kits to extract RNA for future use.

[0039] (2) B.1.1.7 mutant strain S gene: with two mutation sites HV69-70del and N501Y, and other sites are wild-type DNA fragments; purchased from a biological company, diluted and dissolved with TE, and used for later use .

[0040] (3) S gene of B.1.351 mutant strain: It contains two mutation sites K417N and N501Y, and the other si...

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Abstract

The invention belongs to the field of molecular detection, and particularly relates to a digital PCR (polymerase chain reaction) detection kit for the novel coronavirus and variants thereof. Firstly, the invention discloses a primer probe composition for simultaneously detecting the novel coronavirus and the variants thereof, wherein the novel coronavirus comprises a wild type, a B.1.1.7 variant, a B.1.351 variant and a P.1 variant; the primer probe composition is designed according to the N gene, the ORF1ab gene and the mutation types of HV69-70del, K417N, K417T and N501Y of the S gene. The invention develops a novel multiple digital PCR system and a kit for detecting the novel coronavirus and the variants thereof by using the PCR system. The multiple digital PCR system solves the problems that different novel coronavirus variants in the prior art need to be separately detected for several times, and the process is tedious.

Description

technical field [0001] The invention belongs to the field of molecular detection, in particular to a digital PCR detection kit for novel coronavirus and its mutant strains. Background technique [0002] The novel coronavirus (SARS-CoV-2) is a single-stranded positive-sense RNA virus that is prone to mutation. Clinically, the symptoms of patients infected with the new coronavirus vary widely, ranging from asymptomatic to critically ill. In addition to differences in individual factors, virus variation may also be an important factor leading to large differences in symptoms among infected patients. Studies have shown that the mutation rate of the new coronavirus is about 2 to 4 mutations per month, and more than 25 mutant strains have been found in known strains. The B.1.1.7 new crown variant, the B.1.351 new crown variant and the P.1 new crown variant have been discovered in many countries. They have mutations in the receptor binding domain (RBD) of the spike protein on the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/686C12N15/11
CPCC12Q1/701C12Q1/686C12Q2563/159C12Q2537/143Y02A50/30
Inventor 朱海涛夏江
Owner 领航医学科技(深圳)有限公司
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