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RT-PCR discriminating diagnosis primers for type 1 and new serotype duck hepatitis virus

A technology of RT-PCR and duck hepatitis virus, which is applied in the field of primers for differential diagnosis of RT-PCR, can solve the problems of high operator requirements and cumbersome optimization of double PCR conditions, and achieve simple identification methods, rapid differential diagnosis detection, fast and accurate, The effect of filling research gaps

Inactive Publication Date: 2016-10-12
INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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Problems solved by technology

However, the relevant methods all need to design specific primers for the specific nucleotide sequences of duck hepatitis virus type 1 and new serotype duck hepatitis virus, and establish a double PCR differential diagnosis method, but the optimization of double PCR conditions is cumbersome and requires more operators. high

Method used

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  • RT-PCR discriminating diagnosis primers for type 1 and new serotype duck hepatitis virus
  • RT-PCR discriminating diagnosis primers for type 1 and new serotype duck hepatitis virus
  • RT-PCR discriminating diagnosis primers for type 1 and new serotype duck hepatitis virus

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Embodiment 1

[0038] 1 Materials and methods:

[0039] 1.1 Experimental reagents and consumables: Viral nucleic acid RNA extraction kit (EasyPure Viral DNA / RNA Kit), RT-PCR kit (EasyScript One-Step RT-PCR SuperMix) were purchased from Beijing Quanshijin Biotechnology Co., Ltd., real-time PCR was purchased from From AXYGEN Corporation;

[0040] 1.2 Experimental strains:

[0041] Duck hepatitis virus type 1 (DHV-1) (GenBank No. JX390983) and new serotype duck hepatitis virus (DHV-N) (GenBank No. EU755009), avian influenza virus, avian paramyxovirus type 1, reovirus, poultry Tambusu virus, duck circovirus, duck plague virus and Muscovy duck parvovirus were all isolated, identified and preserved by the Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences.

[0042] 1.3 Method:

[0043] 1.3.1 Primer design for RT-PCR:

[0044] Oligo7 was used to design primers. When designing the primers, both DHV-1 and DHV-N can be amplified simultaneously, and there...

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Abstract

The invention discloses a group of RT-PCR discriminating diagnosis primers for discriminating type 1 and new serotype duck hepatitis virus. A primer sequence is shown as a SEQ ID NO. 1-2. The primer is designed according to existed difference of a 3'-terminal nucleotide sequence of a type 1 and new serotype duck hepatitis virus whole-genome sequence, The group of the primers performs discriminating diagnosis in a specific amplification area for amplifying the type 1 and new serotype duck hepatitis virus according to the length difference of the nucleotide sequences, wherein size of a type 1 duck hepatitis virus amplification fragment is about 258 bp, and the size of a new serotype duck hepatitis virus amplification fragment is about 321 bp. The identification method of the primer has the advantages of easy and rapid operation, and high accuracy, only one group of the primers can discriminate the type 1 and new serotype duck hepatitis virus types, and the primers provide technical guarantee for aquatic bird healthy culture.

Description

technical field [0001] The invention belongs to the technical field of rapid diagnosis of veterinary infectious diseases, and in particular relates to a group of RT-PCR differential diagnosis primers for distinguishing type 1 duck hepatitis virus and new serotype duck hepatitis virus. Background technique [0002] Duck hepatitis virus type 1 (DHV-1) is an acute, highly contagious infectious disease caused by duck hepatitis virus, mainly affecting ducklings under 3 weeks old, with hepatitis as the typical feature. The disease has an acute onset, short course, high morbidity and mortality, and seriously threatens the healthy development of the duck industry [Yang Weixing, Shi Shaohua, Chen Hongmei, et al. Sequence analysis of three strains of duck hepatitis virus type 1 genome[J]. China Agricultural Science Bulletin, 2010, 26 (14): 8-12. ]. [0003] In 1999, Su Jingliang of China Agricultural University and others isolated two new serotypes of duck hepatitis virus (Duck hep...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11C12R1/93
CPCC12Q1/706
Inventor 万春和施少华黄瑜程龙飞傅光华陈红梅傅秋玲陈翠腾刘荣昌
Owner INST OF ANIMAL HUSBANDRY & VETERINARY FUJIAN ACADEMY OF AGRI SCI
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