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
作者： Yin, Jiyuan 1 ; Wang, Qing 1 ; Wang, Yingying 1 ; Li, Yingying 1 ; Zeng, Weiwei 1 ; Wu, Jiexing 1 ; Ren, Yan 1 ; Tang, Ya 1 ;
作者机构： 1.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Aquat Anim Immune Technol Guangdong Prov, Key Lab Fishery Drug Dev,Minist Agr, Guangzhou, Guangdong, Peoples R China
2.Northwest Agr & Forestry Univ, Coll Vet Med, Yangling, Shaanxi, Peoples R China
3.Tianjin Agr Univ, Coll Fisheries, Tianjin, Peoples R China
4.FLI, Fed Res Inst Anim Hlth, Inst Infectol, Greifswald, Germany
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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