Rapid detection of duck hepatitis virus type-1 by reverse transcription loop-mediated isothermal amplification

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• 作者：Cuiping Song ; Hongquan Wan ; Shengqing Yu ; Xiangan Han ; Xusheng Qiu ; Qinghai Hu ; Lei Tan ; Chan Ding ; ^{shoveldeen@shvri.ac.cn}
• 关键词：Duck hepatitis ; Reverse transcription loop-mediated isothermal amplification (RT-LAMP) ; Sensitivity ; Specificity
• 刊名：Journal of Virological Methods
• 出版年：2012
• 出版时间：June, 2012
• 年：2012
• 卷：182
• 期：1-2
• 页码：76-81
• 全文大小：749 K

文摘

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of duck hepatitis virus type-1 (DHV-1) was established. Using primers specific to the highly conserved 3D gene of DHV-1, the developed RT-LAMP assay detected the viral RNA of DHV-1 extracted from both allantoic fluid and liver samples of infected ducks. The assay is as sensitive as RT-PCR, and shows no cross-reaction with other common avian viral and bacterial pathogens. In addition to detection via ethidium bromide staining following gel electrophoresis, naked-eye observation after staining with SYBR Green I dye can be used to detect RT-LAMP products; this enables field application of this assay. The findings demonstrate that RT-LAMP can serve as a helpful tool for the detection and surveillance of DHV-1 in the poultry industry.