鸭副黏病毒和鸭圆环病毒二重荧光定量PCR检测方法的建立

›› 2013, Vol. 40 ›› Issue (4): 26-31.

鸭副黏病毒和鸭圆环病毒二重荧光定量PCR检测方法的建立

许宗丽^1,2, 谢芝勋², 谢丽基², 刘加波², 谢志勤², 庞耀珊², 邓显文², 范晴²

1. 广西大学,动物科学技术学院,广西南宁 530004;
2. 广西壮族自治区兽医研究所, 广西畜禽疫苗新技术重点实验室,广西南宁 530001

收稿日期:2012-09-06 出版日期:2013-04-20 发布日期:2013-04-19
通讯作者: 谢芝勋,研究员, 主要从事动物传染病病原分子生物学研究。E-mail: xiezhixun@126.com;Tel:0771-3105702 E-mail:xiezhixun@126.com
作者简介:许宗丽(1984-),女,广西人,硕士,从事动物传染病病原分子生物学研究。
基金资助:
广西科技项目(桂科攻10100014-5、桂科重1222003-2-4、桂科专项11-3);广西特聘专家专项经费(2011B020);广西重点实验室建设项目(12-071-28)共同资助。

Establishment of a Duplex Real-time PCR Assay for Detection of Duck Paramyxovirus and Duck Circovirus

XU Zong-li^1,2, XIE Zhi-xun², XIE Li-ji², LIU Jia-bo², XIE Zhi-qin², PANG Yao-shan², DENG Xian-wen², FAN Qing²

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

Received:2012-09-06 Online:2013-04-20 Published:2013-04-19

摘要/Abstract

摘要： 根据鸭副黏病毒(DPMV)和鸭圆环病毒(DuCV)保守基因序列,设计了2对针对鸭副黏病毒和鸭圆环病毒的特异性引物和2条不同荧光基团标记的TaqMan探针,建立了鸭副黏病毒和鸭圆环病毒的二重荧光定量PCR检测方法。该方法敏感性好,对鸭副黏病毒和鸭圆环病毒的检测敏感性分别达到160和140个拷贝数;该方法特异性强,对鸭肝炎病毒、番鸭细小病毒、鸭瘟病毒和H9型禽流感病毒等病原体的检测全为阴性;应用该方法对118份临床病料进行检测,结果检出鸭副黏病毒和鸭圆环病毒阳性感染率分别为0.85%和8.47%,无混合感染。本试验建立的二重荧光定量PCR具有快速、特异、敏感和重复性好等优点,适用于鸭副黏病毒和鸭圆环病毒的快速诊断和监测。

关键词: 鸭副黏病毒; 鸭圆环病毒; 二重荧光定量PCR

Abstract: A duplex Real-time polymerase chain reaction(drRT-PCR) assay was developed and optimized to simultaneously detect duck paramyxovirus and duck circovirus in one reaction. Two sets of specific primers for duck paramyxovirus and duck circovirus, along with two TaqMan probes specific for each virus were used in the assay. This drRT-PCR assay was found to be specific and be able to detect and differentiate duck paramyxovirus and duck circovirus, and no positive results were observed, when nucleic acid from duck hepatitis virus, Muscovy duck parvovirus, duck plague virus and H9 subtype avian influenza virus and so on were used as drRT-PCR templates. The sensitivity of this drRT-PCR assay was 160 and 140 copies for duck paramyxovirus and duck circovirus,respectivly. The drRT-PCR assay was applied to 118 clinical samples detection. Among these samples, duck paramyxovirus positive rate was 0.85%, duck circovirus positive tate was 8.47%, no co-infection. This drRT-PCR assay was a quick, sensitive and specific test for detecting duck paramyxovirus and duck circovirus,and would be useful for the control of these viruses in ducks.

Key words: duck paramyxovirus; duck circovirus; duplex Real-time PCR

中图分类号:

许宗丽, 谢芝勋, 谢丽基, 刘加波, 谢志勤, 庞耀珊, 邓显文, 范晴. 鸭副黏病毒和鸭圆环病毒二重荧光定量PCR检测方法的建立[J]. , 2013, 40(4): 26-31.

XU Zong-li, XIE Zhi-xun, XIE Li-ji, LIU Jia-bo, XIE Zhi-qin, PANG Yao-shan, DENG Xian-wen, FAN Qing. Establishment of a Duplex Real-time PCR Assay for Detection of Duck Paramyxovirus and Duck Circovirus[J]. , 2013, 40(4): 26-31.

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