Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of avian reovirus.
Avian reovirus (ARV) is an important pathogen of poultry and causes significant economic losses to the poultry industry. To develop a rapid and sensitive method for the surveillance of ARV, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was established using a set of six primers specific to the S1 gene segment of ARV. The established assay was performed at 62°C for 60 min in a thermal block, and the result was visualized directly under daylight or ultraviolet light. The detection limit of the RT-LAMP assay was 10 fg total RNA, which was 100-fold higher than that of reverse transcriptase polymerase chain reactions. The specificity of the assay was supported by the lack of cross-reaction with other avian pathogens. Furthermore, viral RNAs of field isolates were successfully detected by the assay. Overall, the newly established RT-LAMP assay is simple, rapid, sensitive, specific, and can visually detect ARV without the use of any specialized equipment.
Department of Biotechnology, Guangxi Key Laboratory of Animal Vaccines and New Technology, Guangxi Veterinary Research Institute, Nanning Guangxi, PR China. s.xiezhixun@126.com
Zhixun
Z
Xie
Yi
Y
Peng
Sisi
S
Luo
Ying
Y
Wang
Jiabo
J
Liu
Yaoshan
Y
Pang
Xianwen
X
Deng
Zhiqin
Z
Xie
Liji
L
Xie
Qing
Q
Fan
Liqiong
L
Teng
Xiuqing
X
Wang
D000818:Animals; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000662:veterinary*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D030101:Orthoreovirus, Avian / Q000235:genetics*; D011201:Poultry Diseases / Q000175:diagnosis* / Q000821:virology*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D048348:Reverse Transcription / Q000235:genetics; D012680:Sensitivity and Specificity
D017931:DNA Primers