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Ring mediate isothermal amplification kit for diagnosing H7N9 hypotype avian influenza virus and application method thereof

A ring-mediated isothermal and avian influenza virus technology, applied in biochemical equipment and methods, recombinant DNA technology, DNA/RNA fragments, etc., can solve the problems of high experimental cost and long detection time

Inactive Publication Date: 2014-10-15
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional PCR reaction has a certain dependence on temperature, requiring a precise temperature control system, long detection time, and high experimental cost

Method used

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  • Ring mediate isothermal amplification kit for diagnosing H7N9 hypotype avian influenza virus and application method thereof
  • Ring mediate isothermal amplification kit for diagnosing H7N9 hypotype avian influenza virus and application method thereof
  • Ring mediate isothermal amplification kit for diagnosing H7N9 hypotype avian influenza virus and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0027] 1. Test materials and methods

[0028] 1.1 Test material

[0029] The artificially synthesized fragments of the HA and NA genes of the H7N9 subtype avian influenza virus are artificially synthesized with reference to the published sequences on Genbank (the sequence number of the HA gene is KC853766.1, and the sequence number of the NA gene is KC853765.1), and the fragments are connected to the pCR II vector and transformed into competent cells DH5α, which were stored in the form of 30% glycerol solution.

[0030] 1.2 Primer design

[0031] The DNAMAN software was used to compare the HA and NA genes of the H7N9 subtype avian influenza virus, and the fragments with large differences between the HA and NA genes were selected as the target sequence, and the online software PrimerExplorer V4 (http: / / primerexplorer.jp / e / ) Design primer sets for loop-mediated isothermal amplification of HA and NA genes. The HA gene loop-mediated isothermal amplification primer set consists ...

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Abstract

The invention discloses a ring mediate isothermal amplification kit for diagnosing H7N9 hypotype avian influenza virus and an application method thereof. The kit is characterized by comprising a an HA gene, an HA gene mediate isothermal amplification primer mixed liquor, a ring mediate isothermal amplification reaction premixed solution, an HA gene and HA gene in vitro transcription RNA positive control material and negative control material and the kit can be used for quickly diagnosing the H7N9 hypotype avian influenza virus. The application method of the kit is characterized in that the H7N9 hypotype avian influenza virus can be diagnosed timely and quickly through an isothermal amplification fluorescence detection system. The method has the advantages that the specificity and sensitivity are high and a simple, convenient and quick approach is provided for preventing and controlling the H7N9 hypotype avian influenza virus and diagnosing the trend of the H7N9 hypotype avian influenza virus.

Description

technical field [0001] The present invention relates to a set of primers, kits and usage methods for rapid virus diagnosis, in particular to a set of loop-mediated isothermal amplification primers, kits and usage methods for diagnosing H7N9 subtype avian influenza virus, belonging to poultry Influenza virus detection field. Background technique [0002] Avian influenza is an acute and highly fatal infectious disease of poultry caused by type A influenza virus. It happened in Italy in 1878, and then it happened in other European countries, South America, Southeast Asia, the United States and the former Soviet Union. Due to too many variations of avian influenza, it is not suitable for effective detection and control, and the risk of its spread has aroused great concern from the international community. On September 29, 2005, Dr. David Nabarro, an expert from the World Health Organization (WHO), who was appointed by UN Secretary-General Kofi Annan as the UN bird flu coordina...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/70C12Q1/6844C12Q2531/119C12Q2545/101C12Q2537/16
Inventor 邹明强薛强殷宏
Owner CHINESE ACAD OF INSPECTION & QUARANTINE