101. Development of a reverse transcription loop-mediated isothermal amplification assay for visual detection of avian reovirus
- Author
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Fan Qing, Xie Zhiqin, Yi Peng, Ying Wang, Xie Zhixun, Xiuqing Wang, Xie Liji, Luo Sisi, Deng Xianwen, Liu Jiabo, Liqiong Teng, and Pang Yaoshan
- Subjects
genetic structures ,Orthoreovirus, Avian ,Loop-mediated isothermal amplification ,Biology ,Sensitivity and Specificity ,Food Animals ,Ultraviolet light ,Animals ,Pathogen ,Gene ,Reverse Transcription Loop-mediated Isothermal Amplification ,Polymerase ,Poultry Diseases ,DNA Primers ,General Immunology and Microbiology ,Reverse Transcriptase Polymerase Chain Reaction ,Reverse Transcription ,Virology ,Molecular biology ,Reverse transcriptase ,Reoviridae Infections ,Visual detection ,Molecular Diagnostic Techniques ,biology.protein ,Animal Science and Zoology ,Chickens ,Nucleic Acid Amplification Techniques - Abstract
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 62oC 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.
- Published
- 2012