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Avian influenza virus H7N9 sub type fluorescent RT-PCR primer group, probe group, kit and method

An avian influenza virus, RT-PCR technology, applied in the field of avian influenza virus H7N9 subtype fluorescent RT-PCR primer set, can solve the problems of difficult double amplification specificity, sensitivity control, etc., to achieve good repeatability, impurity little effect

Inactive Publication Date: 2018-01-02
奥斯汀生命科学技术公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dual fluorescent RT-PCR, which has completed the initial confirmation of avian influenza virus H7 subtype and N9 subtype in one test, has also been gradually developed, but the difficulty lies in the control of the specificity and sensitivity of the double amplification.

Method used

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  • Avian influenza virus H7N9 sub type fluorescent RT-PCR primer group, probe group, kit and method
  • Avian influenza virus H7N9 sub type fluorescent RT-PCR primer group, probe group, kit and method
  • Avian influenza virus H7N9 sub type fluorescent RT-PCR primer group, probe group, kit and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] A kind of avian influenza virus H7N9 subtype fluorescent RT-PCR primer set, comprises following primers:

[0060] H7-F: 5'-TCACAGCAAATACAGGGAAGAG-3';

[0061] H7-R: 5'-CCCGAAGCTAAACCAGAGTATC-3';

[0062] N9-F: 5'-TAGCAATGACACACACTAGTCAAT-3';

[0063] N9-R: 5'-ATTACCTGGATAAGGGTCATTACACT-3';

[0064] HA-F: 5'-CTTCTAACCGAGGTCGAAACGTA-3';

[0065] HA-R: 5'-GGTGACAGGATTGGTCTTGTCTTTA-3'.

Embodiment 2

[0067] A kind of avian influenza virus H7N9 subtype fluorescent RT-PCR probe set, comprises following probe:

[0068] H7-P5: 5'-ROX-TGACCCAGTCAAACTAAGCAGCGG-MGB-3';

[0069] N9-p7: 5'-JOE-AGACAATCCCCGACCGAATGACCC-MGB-3';

[0070] HA-P2: 5'-CY5-TCAGGCCCCCTCAAAGCCGAG-MGB-3'.

Embodiment 3

[0072] A kind of avian influenza virus H7N9 subtype fluorescent RT-PCR kit, comprises the primer group described in embodiment 1 and the probe group described in embodiment 2.

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Abstract

The invention provides an avian influenza virus H7N9 sub type fluorescent RT-PCR primer group and a probe group matched with the primer group in use. The primer group comprises specific primers and probes designed for H7 antigens and N9 antigens and universal primers and probes for avian influenza virus, a lot of highly specific fragments can be amplified, double RT-PCR amplification is achieved by operation at one time, and the avian influenza virus and H7N9 sub types can be detected according to collecting variations of different fluorescent signals. The invention also provides an avian influenza virus H7N9 sub type fluorescent RT-PCR kit and a corresponding application method. Envelops and nucleocapsid on the outer side of the virus are damaged by means of a nucleic acid oriented release agent to release nucleoid RNA, and a proper liquid environment is provided for the RT-PCR reaction through PCR mixed liquid, so that follow-up detection and analysis are facilitated. The product andthe method provided by the invention have the advantages of sensitivity, specificity, stability, simplicity and convenience, good repeatability and the like, and is suitable for large-scaled antigendetection of front line quarantine officers.

Description

technical field [0001] The invention belongs to the technical field of virus detection, and in particular relates to a fluorescent RT-PCR primer set, probe set, kit and method for avian influenza virus H7N9 subtype. Background technique [0002] Avian influenza virus, which belongs to type A influenza virus, is divided into three levels of high, medium and low / non-pathogenicity according to the pathogenicity of avian influenza virus to chickens and turkeys. Due to the characteristics of the hemagglutinin structure of the avian influenza virus, it usually infects poultry. When the virus undergoes gene reassortment during the replication process, resulting in a change in structure and the ability to infect humans, it may cause human infection with avian influenza. The subtypes of avian influenza virus that can directly infect humans have been found so far: H5N1, H7N1, H7N2, H7N3, H7N7, H9N2 and H7N9 subtypes. Among them, the highly pathogenic H5N1 subtype and the new avian in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
Inventor 许智慧
Owner 奥斯汀生命科学技术公司
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