Fluorescent quantitative primer group for visual differential diagnosis of waterfowl parvoviruses

A waterfowl parvovirus and differential diagnosis technology, applied in the direction of microorganism-based methods, microorganism measurement/testing, microorganisms, etc., to achieve the effect of simple identification method, high efficiency and high accuracy

Active Publication Date: 2015-07-08
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

[0007] At present, there are no relevant research reports at home and abroad that only need a set of real-time fluorescent quantitative PCR primers to simultaneously perform visual differential diagnosis of GPV and MDPV infection. The establishment of the present invention can fill in the gaps in related fields at home and abroad

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  • Fluorescent quantitative primer group for visual differential diagnosis of waterfowl parvoviruses

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Embodiment 1

[0029] 1. Virus strains:

[0030] Goose parvovirus and Muscovy duck parvovirus were isolated, identified and preserved by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0031] 2. Primer design and synthesis

[0032] Real-time fluorescence quantitative PCR primers P1 and P2 were designed according to the characteristics of GPV and MDPV non-structural protein genes, and the sequences of primers P1 and P2 were:

[0033] Upstream primer P1: 5'-TTCTTTGCTGCTCTGTTGGAAATA -3',

[0034] Downstream primer P2: 5'-GCTTTTACCAATATGCC-3'.

[0035] 3. Real-time fluorescent quantitative PCR amplification

[0036] GPV and MDPV genomic DNA were extracted by conventional methods. The designed specific real-time fluorescent quantitative PCR primers P1 and P2 were used for real-time fluorescent quantitative PCR amplification.

[0037] The optimized 20 μL optimal reaction system is the system: SYBR Premix Ex Taq TM 10 μL, 0.2 μL each of ...

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Abstract

The invention discloses a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) primer group for visual detection on infection conditions of goose parvovirus (GPV) and muscovy duck parvovirus (MDPV) and a method thereof. According to the method, the detection on the infection conditions of GPV and MDPV is carried out by using dissolution curve temperature difference caused by the difference between nucleotide GC contents of GPV and MDPV specific gene fragment regions amplified by primers, and the infection conditions of GPV and MDPV can be subjected to visual differential diagnosis specifically by only combining the SYBR Green I based real-time fluorescent quantitative PCR primer group to dissolution curves which are automatically generated after reaction is ended. The method disclosed by the invention is simple and is relatively high in efficiency and accuracy.

Description

technical field [0001] The invention belongs to the field of animal molecular etiology, and in particular relates to a set of fluorescent quantitative primers for visual differential diagnosis of water fowl parvovirus. Background technique [0002] Goose parvovirus (GPV) is the main autonomously replicating parvovirus that infects poultry. It was first discovered in 1956 by Chinese scholar Fang Dingyi in Yangzhou, Jiangsu Province (Fang Dingyi. Introduction to Goose Plague[J]. Chinese Journal of Veterinary Medicine, 1962, 8:19-20.], the virus was also isolated in many countries later, the World Society of Poultry named the disease Derzsy in commemoration of the advanced work of the Hungarian scholar Derzsy ’ S disease (Wan Chunhe, Zhu Haixia, Huang Yu, et al. Genetic analysis of a strain of goose parvovirus [J]. Chinese Journal of Animal Infectious Diseases, 2011, 1, 9 (4): 19-24.). [0003] Goose parvovirus belongs to the family Parvoviridae and the genus Parvovirus, and ...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/686C12Q1/70C12Q2561/113C12Q2527/107C12Q2563/107
Inventor 万春和黄瑜陈红梅施少华傅秋玲傅光华程龙飞
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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