Centex Shrimp, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand.
J Virol Methods. 2011 Oct;177(1):71-4. doi: 10.1016/j.jviromet.2011.06.020. Epub 2011 Jul 5.
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°C 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 1h and did not require electrophoresis. Compared to an existing RT-PCR method, the RT-LAMP-LFD was also ∼1000-fold more sensitive in detecting LSNV RNA.
拉明辛格病毒（LSNV）最近在泰国养殖的巨型虎虾（Penaeus monodon）中被发现，这些虾表现出生长缓慢综合征的迹象。环介导等温扩增（LAMP）允许在恒定温度下快速扩增 DNA。在这里，一种逆转录（RT）-LAMP 方法与色谱横向流动条（LFD）相结合，可快速特异性地检测 LSNV RNA。该反应在 65°C 下优化 30 分钟，扩增的 DNA 与 FITC 标记的寡核苷酸探针杂交 5 分钟后，在 LFD 测试线 5 分钟后即可检测到。包括快速 RNA 提取的 10 分钟，在 1 小时内可以生成测试结果，并且不需要电泳。与现有的 RT-PCR 方法相比，RT-LAMP-LFD 检测 LSNV RNA 的灵敏度也提高了约 1000 倍。
