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

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• 作者：Narong Arunrut^a ; ^b ; Yortyot Seetang-Nun^a ; ^b ; Jurairat Phromjai^a ; Wattana Panphut^a ; Wansika Kiatpathomchai^a ; ^b ; ^{wansika@biotec.or.th}
• 关键词：Laem-Singh virus ; Loop-mediated isothermal amplification ; Lateral flow dipstick ; LAMP ; PCR ; LSNV
• 刊名：Journal of Virological Methods
• 出版年：2011
• 出版时间：October, 2011
• 年：2011
• 卷：177
• 期：1
• 页码：71-74
• 全文大小：1K

文摘

Laem-Singh virus (LSNV) was discovered recently in Thailand in farmed Giant Tiger shrimp (Penaeus monodon) displaying signs of slow growth syndrome. Loop-mediated isothermal amplification (LAMP) allows DNA to be amplified rapidly at a constant temperature. Here a reverse transcription (RT)-LAMP method was combined with a chromatographic lateral-flow dipstick (LFD) to detect LSNV RNA rapidly and specifically. The reaction was optimized at 65 ¡ãC for 30 min and amplified DNA hybridized to an FITC-labeled oligonucleotide probe for 5 min was detected at LFD test line 5 min after application. Including 10 min for rapid RNA extraction, test results could be generated within 1 h and did not require electrophoresis. Compared to an existing RT-PCR method, the RT-LAMP-LFD was also ?000-fold more sensitive in detecting LSNV RNA.