High-sensitivity method used for detecting and identifying human coronavirus

A technology of human coronavirus and kit, applied in biochemical equipment and methods, measurement/testing of microorganisms, resistance to vector-borne diseases, etc., can solve problems such as missed detection

Active Publication Date: 2014-01-01
INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI +1
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Due to certain mutations in the HCoV genome, this method will cause a certain degree of m

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  • High-sensitivity method used for detecting and identifying human coronavirus

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[0042] The present invention includes simultaneous detection of one or more of 6 known HCoVs, for HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV using 2-fold reaction for detection of specific sequence regions, for MERS-CoV was detected using a 4-plex reaction targeting specific sequence regions. The specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0043] According to the method for highly sensitive detection and / or identification of human coronaviruses provided by the embodiments of the present invention, six kinds of human coronaviruses to be detected include: HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS- CoV. Including the following steps:

[0044] (1) According to the whole genome sequence of the HCoV to be tested, select the specific conserved sequence of each HCoV, design PCR primers, and have a tag sequence of any combination of 10 bases (ACGTTGGATG) at the 5' end of the pr...

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Abstract

The invention provides a high-sensitivity method used for detecting and identifying 6 kinds of human coronavirus. The high-sensitivity method is based on exclusion of pollution caused by PCR amplification products, and is used for detecting and/or identifying human coronavirus by taking multiplex PCR-mass spectrometry as a platform. The 6 kinds of human coronavirus include HCoV-229E, HCoV-OC43, HCoV-NL63, HCoV-HKU1, SARS-CoV and MERS-CoV. Detection parts are the specific sequence domains in the genomes of the 6 kinds of human coronavirus. The high-sensitivity method is capable of realizing multiple gene detection of the 6 kinds of human coronavirus in a reaction, and increasing detectable rate; possesses advantages of high sensitivity and specificity, convenient operation, speediness, high throughput and the like; and can be used in scientific researches, clinical and epidemiological investigation.

Description

technical field [0001] The invention relates to a method for detecting pathogenic microorganisms, in particular to a method for detecting and identifying human coronaviruses with high sensitivity. Background technique [0002] Human coronavirus (HCoV) belongs to the Coronavirus family (Coronaviridae) and is a single-stranded positive-sense RNA virus. Virus particles are irregular in shape, surrounded by a fatty membrane, and there are three glycoproteins on the surface of the membrane: spike glycoprotein (S, Spike Protein); small envelope glycoprotein (E, Envelope Protein); membrane glycoprotein (M, Membrane Protein) . [0003] HCoV is one of the main pathogens of the common cold in adults. It can cause upper respiratory tract infection in children, and generally rarely affects the lower respiratory tract. There are six kinds of HCoV that have been discovered so far: HCoV-229E, HCoV-OC43, SARS-CoV, HCoV-NL63, HCoV-HKU1 and MERS-CoV. Among them, the earliest discovered HCo...

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/6848C12Q1/686C12Q1/70C12Q2521/525C12Q2521/531C12Q2537/143C12Q2565/627C12Q2531/113Y02A50/30
Inventor 彭俊平郭军华金奇
Owner INST OF PATHOGEN BIOLOGY CHINESE ACADEMY OF MEDICAL SCI
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