Rapid and sensitive detection of infectious bursal disease virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick

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• 作者：Su-Ming Tsai^a ; Hung-Jen Liu^b ; Jui-Hung Shien^c ; Long-Huw Lee^c ; Po-Chung Chang^a ; Chi-Young Wang^c ; ^{cyoungwang@dragon.nchu.edu.tw}
• 关键词：Infectious bursal disease virus ; Reverse transcription loop-mediated isothermal amplification (RT-LAMP) ; Nested RT-PCR ; Lateral flow dipstick (LFD)
• 刊名：Journal of Virological Methods
• 出版年：2012
• 期刊代码：180_01660934
• 类别：med
• 出版时间：April, 2012
• 卷：181
• 期：1
• 页码：117-124
• 文件大小：1529 K

摘要

Infectious bursal disease (IBD), an immunosuppressive disease that affects all ages of chickens, results in significant losses in the poultry industry. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) combined with a chromatographic lateral flow dipstick (LFD) for the detection of infectious bursal disease virus (IBDV) was developed. The whole process of testing can be completed in less than 70 min using biotin-labeled primers, an FITC-labeled DNA probe, and the LFD. The detection limits for IBDV using RT-LAMP and RT-LAMP-LFD were the same at 10^鈭? plaque forming units (PFU). When other unrelated viruses and cells were tested, no false positive results were observed. In addition, the amplification efficiency of RT-LAMP was enhanced when a loop primer was used. The RT-LAMP-LFD product started to be detected after 40 min. Clinical samples were used to compare assays using RT-PCR, nested RT-PCR, RT-LAMP, and RT-LAMP-LFD and the positive rates were 16%, 40%, 40%, and 40%, respectively. In conclusion, this assay is an easy, rapid, accurate, and sensitive method for the detection of IBDV and will improve the screening of field samples, especially when veterinarians have limited resources.