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鸭圆环病毒与鸭Ⅰ型肝炎病毒二重 PCR 检测方法的建立
引用本文:赵光远,谢芝勋,谢丽基,庞耀珊,谢志勤,邓显文,刘加波,范晴.鸭圆环病毒与鸭Ⅰ型肝炎病毒二重 PCR 检测方法的建立[J].生物技术通讯,2012,0(6):863-865.
作者姓名:赵光远  谢芝勋  谢丽基  庞耀珊  谢志勤  邓显文  刘加波  范晴
作者单位:广西大学动物科技学院;广西兽医研究所广西疫苗新技术重点实验室
基金项目:广西科技项目(桂科攻10100014-5和桂科专项11-3);广西特聘专家专项(2011B020);国家百千万人才工程人选专项(945200603)
摘    要:目的:建立一种同时检测鸭圆环病毒(DuCV)和鸭I型肝炎病毒(DHV)病原体的二重PCR技术。方法:根据DuCV和DHV的基因文库,分别设计了2对与DuCV和DHV某段基因序列互补的引物,用这2对引物对同一样品中DuCV和DHV模板进行二重PCR扩增。结果与结论:用建立的方法均同时得到了2条特异性的大小与实验设计相符(DuCV:245bp;DHV:569bp)的二重PCR扩增带,而且对其他禽病病原的PCR扩增结果均为阴性,能同时检出56Pg的DHVRNA模板和6Pg的DuCVDNA模板。

关 键 词:鸭圆环病毒  鸭Ⅰ型肝炎病毒  二重PCR

Detection of Duck Circovirus and Duck Hepatitis Virus 1 by Multiplex PCR
ZHAO Guang-Yuan,XIE Zhi-Xun,XIE Li-Ji,PANG Yao-Shan,XIE Zhi-Qin,DENG Xian-Wen,LIU Jia-Bo,FAN Qing.Detection of Duck Circovirus and Duck Hepatitis Virus 1 by Multiplex PCR[J].Letters in Biotechnology,2012,0(6):863-865.
Authors:ZHAO Guang-Yuan  XIE Zhi-Xun  XIE Li-Ji  PANG Yao-Shan  XIE Zhi-Qin  DENG Xian-Wen  LIU Jia-Bo  FAN Qing
Institution:1.College of Veterinary Medicine,Guangxi University,Nanning 530004;2.Guangxi Veterinary Research Institute,Guangxi Key Laboratory of Anmial Vaccines and New Technology,Nanning 530001;China
Abstract:Objective: A multiplex PCR was optimized to simuhaneously detect two pathogenic of duck hepatitis virus I(DHV) and duck circovirus(DuCV). Methods: Two sets of specific primers were designed according to the sequences of DHV and DuCV according to the nucleotide sequence in GenBank. A multiplex PCR was carried out with the two specific primers. Results & Conclusion: All the samples which contained DHV cDNA and DuCV DNA, could be amplified by the multiplex PCR using these two sets primers, yielding two specific bands of DHV 569 bp and DuCV 245 bp. No specific band was amplified from other avian pathogenic virus and bacte- ria. It turned out that as little as 56 pg of DHV RNA and 6 pg of DuCV DNA was detected using gel electropho- resis in this multiplex PCR.
Keywords:duck hepatitis virus 1  duck circovirus  multiplex PCR
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