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Multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus, and detection method and application thereof

A technology of goose paramyxovirus and goose parvovirus, applied in the field of virus detection, to achieve good amplification effect and high detection accuracy

Inactive Publication Date: 2019-04-12
POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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Problems solved by technology

However, in the prior art, there are no relevant reports on the detection of goose astrovirus, goose paramyxovirus, and goose parvovirus using multiplex PCR.

Method used

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  • Multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus, and detection method and application thereof
  • Multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus, and detection method and application thereof
  • Multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus, and detection method and application thereof

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Embodiment 1

[0040] 1 Materials and methods

[0041] 1.1 Virus strains goose paramyxovirus (GPMV), goose astrovirus (GoAst), goose parvovirus (GPV), H9 subtype avian influenza virus (H9 AIV), by Poultry Research Institute, Shandong Academy of Agricultural Sciences Room provided. The H5 subtype avian influenza virus (H5AIV) positive plasmid was donated by the Poultry Disease Laboratory, School of Veterinary Medicine, South China Agricultural University.

[0042] 1.2 The main reagents: Spin column virus DNA / RNA extraction kit was purchased from Beijing Quanshijin Biotechnology Co., Ltd.; HiScript cDNA first-strand synthesis kit and Premix rTaq enzyme were purchased from Nanjing Novozan Biotechnology Co., Ltd.; DL2000 DNA Marker was purchased from Intel Biotech Co., Ltd.

[0043] 1.3 Test method

[0044] 1.3.1 Design and synthesis of primers Design 3 pairs of specific primers that can amplify 3 kinds of goose pathogens and store them at -20℃ for later use. The sequences of the 3 pairs of primers a...

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Abstract

The invention relates to the technical field of virus detection, and specifically relates to multiplex PCR detection primer pairs for goose astrovirus, goose paramyxovirus and goose parvovirus; and the invention further relates to a detection method of the primer pairs and application of the primer pairs. The multiplex PCR system established by the invention is capable of accurately detecting simplex or mixed infection of waterfowl with the goose astrovirus, the goose paramyxovirus and the goose parvovirus. Moreover, the multiplex PCR system is optimized with determined optimal primer ratio, primer concentration and Premix rTaq enzyme; and thus, better DNA amplification effects of the 3 viruses are achieved. Therefore, the multiplex PCR system has higher detection accuracy, and thereby having important advantages in the field of virus identification.

Description

Technical field [0001] The invention relates to the technical field of virus detection, in particular to a primer pair for multiple PCR detection of goose astrovirus, goose paramyxovirus, and goose parvovirus, as well as a detection method using the primer pair and the application of the primer pair. Background technique [0002] Astroviruses belong to the astrovirus family (astroviridae), and are divided into two genera according to the host: Mamastrovirus and Avastrovirus. There are 3 genera of Avian Astrovirus (Avastrovirus 1-3), including Duck astrovirus, Avian nephritisa strovirus, Turkey astrovirus, etc. Shape virus is its representative species. Astrovirus is a single-stranded, non-enveloped small RNA virus. Its genome is about 6.8kb in length. It contains 4 parts from 5'end to 3'end: one 85-nucleotide 5'end non- Coding region, 3 open reading frames (ORFs) (ORF1a, ORF1b, ORF2), 1 80-nucleotide 3'non-coding region, 1 30-nucleotide polyadenylic acid tail. Since 2017, the...

Claims

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Application Information

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IPC IPC(8): C12Q1/70C12Q1/686C12N15/11C12R1/93
CPCC12Q1/686C12Q1/701C12Q2600/16C12Q2537/143
Inventor 孟凯王友令袁小远张玉霞杨金星
Owner POULTRY INST SHANDONG ACADEMY OF AGRI SCI
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