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

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          Abstract

          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°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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          Author and article information

          Journal
          J Fish Dis
          Journal of fish diseases
          Wiley
          1365-2761
          0140-7775
          Jun 2019
          : 42
          : 6
          Affiliations
          [1 ] Key Laboratory of Fishery Drug Development of Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.
          [2 ] College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling, China.
          [3 ] College of Fisheries, Tianjin Agriculture University, Tianjin, China.
          [4 ] Institute of Infectology, Friedrich-Loeffler-Institut (FLI), Federal Research Institute for Animal Health, Greifswald-Insel Riems, Germany.
          Article
          10.1111/jfd.12983
          30920677
          7d92952e-bde7-4f36-bf61-86bcc41430f4
          © 2019 John Wiley & Sons Ltd.

          PCR,detection,loopmediated isothermal amplification,tilapia lake virus

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