Development of a simple and rapid reverse transcription-loopmediated isothermal amplification (RT-LAMP) assay for sensitive detection of tilapia lake virus

文献类型： 外文期刊

第一作者： Yin, Jiyuan

作者： Yin, Jiyuan;Wang, Qing;Wang, Yingying;Li, Yingying;Zeng, Weiwei;Wu, Jiexing;Ren, Yan;Tang, Yafang;Gao, Caixia;Hu, Huzi;Tang, Yafang;Gao, Caixia;Hu, Huzi;Bergmann, Sven M.

作者机构：

关键词： detection; loopmediated isothermal amplification; PCR; tilapia lake virus

期刊名称：JOURNAL OF FISH DISEASES （ 影响因子：2.767； 五年影响因子：2.689 ）

ISSN： 0140-7775

年卷期： 2019 年 42 卷 6 期

页码：

收录情况： SCI

摘要： Recently, substantial mortality of farmed and wild tilapia caused by tilapia lake virus (TiLV) infection has been observed worldwide. However, sensitive and reliable diagnostic method is limited. A reverse transcription-loopmediated isothermal amplification (RT-LAMP) assay has been applied for the detection of TiLV nucleotide sequence. Six primers targeting two locations on the target gene based on a highly conserved sequence in the segment 1 (S1) region of the TiLV genome have been designed. The optimized RT-LAMP reaction was maintained at the isothermal condition of 63 degrees C for 45 min. And the amplifications could be verified by turbidity or a colour change with the addition of SYBR Green I. Subsequently, RT-LAMP products could be observed by a ladder pattern following gel electrophoresis. The species-specific assay showed that the method was sensitive enough to detect as low as 1.6 copies of viral particle, and the assay was highly specific because no cross-reactivity was observed with other pathogens, and had a diagnostic sensitivity and specificity of 100% when TiLV-positive samples and non-target virus were tested. In summary, all the results demonstrate that this RT-LAMP is a rapid, effective and sensitive method for TiLV detection in tilapia aquaculture.

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