禽流感病毒与鸡细小病毒二重PCR检测方法的建立

doi:10.16431/j.cnki.1671-7236.2017.07.031

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (7): 2126-2131.doi: 10.16431/j.cnki.1671-7236.2017.07.031

禽流感病毒与鸡细小病毒二重PCR检测方法的建立

何莹^1,2, 谢芝勋², 罗思思², 李孟², 谢丽基², 邓显文², 谢志勤², 奉彬², 黄娇玲², 张艳芳²

1. 广西大学动物科学技术学院, 南宁 530004;
2. 广西壮族自治区兽医研究所, 广西兽医生物技术重点实验室, 南宁 530001

收稿日期:2017-01-05 出版日期:2017-07-20 发布日期:2017-07-22
通讯作者: 谢芝勋 E-mail:xiezhixun@126.com
作者简介:何莹(1990-),女,广西北流人,硕士生,研究方向:禽病防治与病原分子生物学,E-mail:heying1101@163.com
基金资助:
广西科技基地和人才专项项目（桂科AD16380009）；桂渔牧科（201204930、201452001）；国家"万人计划"领军人才专项（2016-37-88）；广西科技项目（15-140-36-A-1）

Development of a Duplex PCR Assay for Detection of Avian Influenza Virus and Chicken Parvovirus

HE Ying^1,2, XIE Zhi-xun², LUO Si-si², LI Meng², XIE Li-ji², DENG Xian-wen², XIE Zhi-qin², FENG Bin², HUANG Jiao-ling², ZHANG Yan-fang²

1. College of Animal Science and Technology, Guangxi University, Nanning 530004, China;
2. Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute Nanning 530001, China

Received:2017-01-05 Online:2017-07-20 Published:2017-07-22

摘要/Abstract

摘要：

为建立一种能同时鉴别诊断禽流感病毒（avian influenza virus，AIV）和鸡细小病毒（chicken parvovirus，ChPV）的检测方法，本研究根据GenBank中AIV的M基因和ChPV的NS基因保守序列，分别设计并筛选出两对特异性引物，用于AIV和ChPV的检测。通过优化反应条件，建立了AIV和ChPV二重PCR检测方法。试验结果表明，该方法特异好，能同时检测AIV和ChPV，对其他常见的禽病病原体均未反应；该法对AIV和ChPV的检测下限均为100 fg；对159份临床样品检测结果与PCR阳性产物测序结果一致。本研究建立的AIV和ChPV二重PCR检测方法具有特异性好、灵敏度高的特点，对AIV和ChPV的防制具有重要意义。

关键词: 禽流感病毒(AIV); 鸡细小病毒(ChPV); 二重PCR; 检测

Abstract:

In order to establish a method to simultaneously detect avian influenza virus (AIV) and chicken parvovirus (ChPV),two pairs of specific primers were designed according to the sequences of AIV M gene and ChPV NS gene in GenBank. The duplex PCR assay was established by optimizing the reaction conditions.The tests showed that this method had high specificity, could simultaneously detect AIV and ChPV and no specific band was amplified for other subtypes avian pathogenic virus. The sensitivity result showed that the lower detection limit of this method was 100 fg. The results of 159 clinical samples were consistent with the sequencing results of PCR positive product. The double PCR methods for detection of AIV and ChPV established in this study had the characteristics of good specificity and high sensitivity, which was of great significance to the prevention control of AIV and ChPV.

Key words: avian influenza virus (AIV); chicken parvovirus (ChPV); duplex PCR; detection

中图分类号:

S852

何莹, 谢芝勋, 罗思思, 李孟, 谢丽基, 邓显文, 谢志勤, 奉彬, 黄娇玲, 张艳芳. 禽流感病毒与鸡细小病毒二重PCR检测方法的建立[J]. 《中国畜牧兽医》, 2017, 44(7): 2126-2131.

HE Ying, XIE Zhi-xun, LUO Si-si, LI Meng, XIE Li-ji, DENG Xian-wen, XIE Zhi-qin, FENG Bin, HUANG Jiao-ling, ZHANG Yan-fang. Development of a Duplex PCR Assay for Detection of Avian Influenza Virus and Chicken Parvovirus[J]. , 2017, 44(7): 2126-2131.

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禽流感病毒与鸡细小病毒二重PCR检测方法的建立

Development of a Duplex PCR Assay for Detection of Avian Influenza Virus and Chicken Parvovirus

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