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Detecting kit and method for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43

A technology of human coronavirus and human metapneumovirus, applied in biochemical equipment and methods, measurement/inspection of microorganisms, resistance to vector-borne diseases, etc., can solve problems such as false positives, long amplification time, laboratory contamination, etc. , to achieve the effect of reducing the generation of primer dimers, improving amplification efficiency, and improving reaction efficiency

Inactive Publication Date: 2014-09-24
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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Problems solved by technology

However, Tem-PCR still has some deficiencies that need to be improved. For example, the sensitivity is slightly lower than that of real-time PCR, and the amplification time is longer than that of traditional PCR. Gene detection and amplification have not yet been integrated, etc.
Therefore, in the detection of respiratory pathogens such as influenza A virus, influenza B virus, human metapneumovirus NL63, OC43 and HKU1 coronavirus, there is no direct, rapid and accurate method for detection at one time.

Method used

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  • Detecting kit and method for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43
  • Detecting kit and method for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43
  • Detecting kit and method for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43

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Embodiment 1

[0122] The present invention detects type A and type B influenza virus, human metapneumovirus, and the kit of NL63 type, HKU1 type and OC43 type human coronavirus, comprising

[0123] Positive control: Escherichia coli with gene fragments of type A influenza virus, Escherichia coli with gene fragments of influenza B virus, Escherichia coli with gene fragments of metapneumovirus HMPV, Escherichia coli with gene fragments of coronavirus NL63, Escherichia coli with coronavirus HKU1 gene fragment, Escherichia coli with coronavirus OC43 gene fragment.

[0124] According to the RT-PCR primers specifically designed for influenza A and B, human metapneumovirus, and NL63, HKU1 and OC43 human coronaviruses, refer to the above, and will not repeat them again:

[0125] And gene probes designed specifically for target genes of influenza A and B, human metapneumovirus, and NL63, HKU1, and OC43 human coronaviruses.

[0126] Also includes the universal primers required in multiplex fluoresce...

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Abstract

The invention provides a detecting kit for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43. The detecting kit comprises a specific pathogen primer and a pathogen gene probe. The detecting kit for influenza viruses A and B, human metapneumovirus and human coronaviruses NL63, HKU1 and OC43 is used for carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) by using the specifically designed pathogen primer and pathogen gene probe on the basis of a multi-fluorescent real-time PCR and Tem-PCR technology, so that the amplification efficiency can be greatly increased to ensure that the detected result can still have very high detecting sensitivity by using a common TaqDNA polymerase, and furthermore, the problems of low amplification efficiency and low detecting sensitivity of the traditional multi-PCR are solved.

Description

technical field [0001] The invention belongs to the technical field of virus detection kits, in particular to a kit and a detection method for detecting type A and type B influenza viruses, human metapneumovirus, NL63 type, HKU1 type and OC43 type human coronavirus. Background technique [0002] Influenza virus is an enveloped, negative-sense RNA virus belonging to the family Orthomyxoviridae. Influenza viruses can be divided into three types: A, B, and C according to the antigenicity of their nucleoproteins and matrix proteins. Currently, types A and B mainly infect humans. The A and B influenza virus genomes are divided into 8 segments, which are single-stranded and negative-strand RNAs, respectively encoding at least 10 kinds of proteins. Type A influenza viruses can be divided into 1-16 HA subtypes and 1-9 NA subtypes according to the antigenicity of their surface cytohemagglutinin (HA) and neuraminidase (NA). Type A influenza virus can infect a variety of animals, suc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
CPCC12Q1/686C12Q1/70C12Q2531/113C12Q2537/143C12Q2563/107C12Q2525/301Y02A50/30
Inventor 房师松张然张仁利
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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