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Primer and probe for real-time fluorescence quantification PCR detection of goose polyoma virus

A real-time fluorescence quantitative, polyoma virus technology, applied in the field of animal infectious diseases, to achieve the effect of rapid detection, good repeatability and strong specificity

Pending Publication Date: 2017-12-26
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, there are currently no relevant research reports on primers, probes and methods for detecting goose polyomavirus based on the TaqMan probe method. The establishment of the present invention can fill in the gaps in related fields at home and abroad.

Method used

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  • Primer and probe for real-time fluorescence quantification PCR detection of goose polyoma virus
  • Primer and probe for real-time fluorescence quantification PCR detection of goose polyoma virus
  • Primer and probe for real-time fluorescence quantification PCR detection of goose polyoma virus

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Embodiment 1

[0031] 1. Design and synthesis of primers and probes

[0032] Referring to the genome sequence of goose polyomavirus in the GenBank database, the molecular biology analysis software Lasergene was used for comparative analysis to clarify the characteristics of the genome sequence, and specific primers (GHPY-F and GHPY-R) and probe (GHPY-P), the relevant primer sequences are:

[0033] Upstream primer GHPY-F: 5'- GCTTCAGATGATGATATGG -3',

[0034] Downstream primer GHPY-R: 5'- CACCCGGAACAAATATTAC -3';

[0035] The probe sequence GHPY-P is: 5'-AAGGATCTTGCACTTGCACGTTC-3', its 5'-end is labeled with a fluorescent reporter group FAM, and its 3'-end is labeled with a fluorescent quencher group Eclipse. Primers and probes were synthesized by Baoriyi Biotechnology (Beijing) Co., Ltd.

[0036] 1. Construction of standard products

[0037] According to the genome characteristics of goose polyomavirus, specific primers were designed by using the primer design software Oligo (version v7....

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Abstract

The invention relates to a primer and a probe for real-time fluorescence quantification PCR detection of goose polyoma virus and belongs to the field of epizootiology. The invention includes design for specific primer and probe sequence, construction for standard plasmid, establishment for real-time fluorescence quantification PCR amplification method, condition optimization, detection result judgment and application. The method for real-time fluorescence quantification PCR detection of goose polyoma virus established by the invention has high sensitivity, high stability, high specificity and high repeatability for detecting the goose polyoma virus, can be used for detecting at least 35.4 copies and can be used for detecting the infection of the goose polyoma virus in a clinic sample.

Description

technical field [0001] The invention relates to a primer and a probe for real-time fluorescent quantitative PCR detection of goose polyomavirus, belonging to the field of zoonotic diseases. Background technique [0002] Goose hemorrhagic polyomavirus (GHPV) can cause goose hemorrhagic nephritis and enteritis of goose (HNEG) in geese, mainly infecting 4-10 week old geese. The main lesions of geese infected with GHPV are hemorrhagic enteritis, hemorrhagic nephritis, ascites and edema. The disease was first reported in 1969, and then it was found to exist in geese flocks in many places in Europe. [0003] GHPY is a non-enveloped circular DNA virus with a total gene length of about 5.2 kb, encoding 5 main open reading frames (open reading frame, ORF): 3 hypothetical proteins (VP1, VP2 and VP3) and two T antigens Proteins (large T antigen and small T antigen). It is generally believed that ducks are not sensitive to GHPY infection; however, it was first reported in 2008 that GH...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/6851C12Q1/701C12Q2531/113C12Q2561/101C12Q2545/113
Inventor 万春和黄瑜程龙飞陈翠腾傅光华施少华陈红梅傅秋玲刘荣昌
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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