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Báo cáo khoa học: "Detection of anatid herpesvirus 1 gC gene by TaqMan™ fluorescent quantitative real-time PCR with specific primers and probe"

Detection of anatid herpesvirus 1 gC gene by TaqMan™ fluorescent quantitative real-time PCR with specific primers and probe | Zou et al. Virology Journal 2010 7 37 http www.virologyj.eom content 7 1 37 VIROLOGY JOURNAL RESEARCH Open Access Detection of anatid herpesvirus 1 gC gene by TaqMan fluorescent quantitative real-time PCR with specific primers and probe Qing ZouH Kunfeng Sun13 Anchun Cheng1 2 Mingshu Wang1 2 Chao Xu1 Dekang Zhu2 Renyong Jia2 Qihui Luo2 Yi Zhou3 Zhengli Chen2 Xiaoyue Chen1 2 3 Abstract Background Anatid herpesvirus 1 AHV-1 is known for the difficulty of monitoring and controlling because it has a long period of asymptomatic carrier state in waterfowls. Furthermore as a significant essential agent for viral attachment release stability and virulence gC UL44 gene and its protein product glycoprotein C may play a key role in the epidemiological screening. The objectives of this study were to rapidly sensitively quantitatively detect gC gene of AHV-1 and provide the underlying basis for further investigating pcDNA3.1-gC DNA vaccine in infected ducks by TaqMan fluorescent quantitative real-time PCR assay FQ-PCR with pcDNA3.1-gC plasmid. Results The repeatable and reproducible quantitative assay was established by the standard curve with a wide dynamic range eight logarithmic units of concentration and very good correlation values 1.000 . This protocol was able to detect as little as 1.0 X 101 DNA copies per reaction and it was highly specific to AHV-1. The TaqMan FQ-PCR assay successfully detected the gC gene in tissue samples from pcDNA3.1-gC and AHV-1 attenuated vaccine AHV-1 Cha strain inoculated ducks respectively. Conclusions The assay offers an attractive method for the detection of AHV-1 the investigation of distribution pattern of AHV-1 in vivo and molecular epidemiological screening. Meanwhile this method could expedite related AHV-1 and gC DNA vaccine research. Background Anatid herpesvirus 1 AHV-1 infection alternatively known as duck virus enteritis DVE or duck plague DP is one of the most widespread and devastating diseases of waterfowls in the family .