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Novel coronavirus fluorescent PCR (Polymerase Chain Reaction) detection kit and use method thereof

A detection kit and coronavirus technology, applied in the biological field, can solve the problems affecting the sensitivity of detection and analysis, the reproducibility of specific detection, low sensitivity and accuracy, so as to improve the situation of DNA denaturation and renaturation, and improve the reaction efficiency and good specificity and reproducibility

Pending Publication Date: 2022-03-29
WUHAN BIOTECH GENE ENG +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the sensitivity and accuracy of fluorescent PCR detection kits for the new coronavirus on the market are still low, and other components of the kit, reaction system, and the amount of samples added will affect the sensitivity, specificity, and reproducibility of detection analysis

Method used

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  • Novel coronavirus fluorescent PCR (Polymerase Chain Reaction) detection kit and use method thereof
  • Novel coronavirus fluorescent PCR (Polymerase Chain Reaction) detection kit and use method thereof
  • Novel coronavirus fluorescent PCR (Polymerase Chain Reaction) detection kit and use method thereof

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Embodiment Construction

[0027] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the implementation manners in the present invention, all other implementation manners obtained by persons of ordinary skill in the art without making creative efforts belong to the scope of protection of the present invention.

[0028] SARS-CoV-2 primers and probes include primers and probes for the new coronavirus ORFlab gene, N gene, and internal standard RNase P gene. details as follows:

[0029] The primers and probe sequences of the new coronavirus ORFlab gene are:

[0030] Forward primer: 5'-ATGCTCGAACTGCACCT-3' (SEQ ID NO: 1);

[0031] Reverse primer: 5'-TAAGCCACTGGTATTTCGC-3' (SEQ ID NO: 2);

[0032] Probe: 5'-TGAGGGACAAGGACACCAAGTGTCTCACCA-3' (SEQ ID NO: 3).

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Abstract

The invention provides a novel coronavirus fluorescent PCR detection kit and a use method thereof. The novel coronavirus fluorescent PCR detection kit comprises primers, probes, Taq DNA polymerase, UNG enzyme, dNTPs, Tris-HCl, MgSO4, glycerol and gelatin, wherein the primers and the probes are used for detecting a novel coronavirus ORFlab gene and an N gene and an internal standard virus RNase P gene; the probes of the ORFlab gene, the N gene and the RNase P gene are all probes modified by locked nucleic acid. The detection limit of the kit is 200 copies / mL, the locked nucleic acid probe, UNG enzyme, glycerol and gelatin are adopted, false negative results can be effectively avoided, and the kit has the advantages of being good in specificity, high in sensitivity and good in reproducibility.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a novel coronavirus fluorescent PCR detection kit and a method for using the same. Background technique [0002] New Coronary Pneumonia (COVID-19) is a lung inflammation caused by the infection of the new coronavirus SARS-CoV-2, which can cause respiratory tract infection in humans, and the population is generally susceptible. Coronaviruses are a large virus family found in animals and humans, and were first isolated from chicken embryos. Prior to this, there were 6 kinds of coronaviruses known to infect humans, of which 4 low-risk types HCoV-229E, HCoV-OC43, HCoV-NL63 and HCoV-HKU1 could only cause common cold symptoms, while high-risk SARS -CoV and MERS-CoV would cause respiratory diseases of global concern. The new coronavirus SARS-CoV-2 is a new member of the genus Coronavirus β, which is a single-stranded RNA virus. After sequencing, its genome sequence is about 30Kb in length...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/701C12Q1/686C12Q2600/166C12Q2563/107C12Q2525/117C12Q2527/125C12Q2545/101
Inventor 熊慧高峰英肖樊
Owner WUHAN BIOTECH GENE ENG
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