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Triple fluorogenic quantitative PCR detection primer, kit and method of DTMUV, EDSV and H9 subtype AIV

A technology for detection kits and detection primers, which are applied in the directions of microorganism-based methods, biochemical equipment and methods, and the determination/inspection of microorganisms to achieve the effect of good repeatability and good amplification effect.

Active Publication Date: 2015-09-09
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

So far, no kits have been reported for simultaneous detection of duck Tembusu virus, egg drop syndrome virus and H9 subtype avian influenza virus

Method used

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  • Triple fluorogenic quantitative PCR detection primer, kit and method of DTMUV, EDSV and H9 subtype AIV
  • Triple fluorogenic quantitative PCR detection primer, kit and method of DTMUV, EDSV and H9 subtype AIV
  • Triple fluorogenic quantitative PCR detection primer, kit and method of DTMUV, EDSV and H9 subtype AIV

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Embodiment Construction

[0067] The present invention will be further described below in conjunction with the examples, but not limited thereto.

[0068] 1 Materials and methods

[0069] 1.1 Viruses Duck Tembusu virus JM strain, egg drop syndrome virus and H9 subtype avian influenza virus are preserved by our laboratory.

[0070] 1.2 Reagents One Step SYBR PrimeScript RT-PCR Kit II, DNA Molecular Weight Standards, pMD18-T Vector, Takara MiniBEST DNA / RNA Extraction Kit, Gel Recovery Kit were purchased from Dalian Bao Biological Engineering Co., Ltd. DH5α Competent was purchased from Tiangen Company, All other reagents were analytically pure.

[0071] 1.3 Primer design and synthesis Download the complete gene sequence of duck Tembusu virus, egg drop syndrome virus and H9 subtype avian influenza virus gene sequence in GenBank, and use Oligo 6.0 to design specific primers for their conserved regions. The primers were synthesized by Invitrogen, and the sequences are shown in Table 1.

[0072] T...

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Abstract

The invention discloses a triple fluorogenic quantitative PCR detection primer, kit and method of the duck tembusu virus disease, the egg drop syndrome virus and the H9 subtype avian influenza virus. The detection primer for the duck tembusu virus disease, the egg drop syndrome virus and the H9 subtype avian influenza virus is shown in SEQ ID NO.1-6. The detection primer and kit can detect the EDSV, the H9 AIV and the DTMUV at the same time. No cross reaction with duck plague virus, goose parovovirus, muscovy duck parvovirus and escherichia coli genomic DNA and duck hepatitis A virus 1, duck hepatitis A virus 3, muscovy duck reovirus and newcastle disease virus RNA exists. The lower limits of detection for EDSV nucleic acid, H9 AIV nucleic acid and DTMUV nucleic acid are 8.0 copies, 4.8 copies and 1.3 copies respectively. The detection primer, kit and method can be used for qualitative and quantitative detection for the EDSV, the H9 AIV and the DTMUV.

Description

technical field [0001] The invention belongs to the field of molecular detection, and in particular relates to a triple fluorescent quantitative PCR detection primer, kit and method for duck Tembusu virus, egg drop syndrome virus and H9 subtype avian influenza virus. Background technique [0002] Duck Tembusu Virus Disease (Duck Tembusu Virus Disease) is an acute infectious disease caused by Duck Tembusu Virus (DTMUV). The disease can cause laying ducks and breeding geese to show obvious symptoms of egg production decline in a short period of time, and the course of the disease can last for several weeks. Recently, there have also been reports of infection in laying hens and meat ducks. The causative agent of the disease belongs to the family Flaviviridae, the genus Flavivirus, and Tembusu virus. Although the mortality rate of the disease is not high, the course of the disease is long and the potential harm is huge. The duck Tembusu virus genome is about 11 kb long, and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
Inventor 孙敏华董嘉文李林林袁建丰邝瑞欢张建峰刘志成张春红
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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