Development of a loop-mediated isothermal amplification assay for rapid detection of iridovirus in the Chinese giant salamander

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• 作者：Yan Meng ; Hui Zhang ; Hongwei Liang ; Lingbing Zeng ; Hanbing Xiao ; Congxin Xie
• 关键词：Chinese giant salamander (Andrias davidianus) ; Iridovirus ; Loop-mediated isothermal amplification (LAMP) ; Molecular detection
• 刊名：Journal of Virological Methods
• 出版年：December, 2013
• 年：2013
• 卷：194
• 期：1-2
• 页码：211-216
• 全文大小：1223 K

文摘

The Chinese giant salamander (Andrias davidianus) iridovirus (GSIV) is an emerging infectious pathogen responsible for severe hemorrhagic disease and high mortality in cultured Chinese giant salamanders. A loop-mediated isothermal amplification (LAMP) assay based on the major caspid protein (MCP) gene has been developed to detect this virus. Primer pairs for the LAMP assay were designed based on the GSIV MCP gene sequence. Amplification results indicate that under optimized conditions the LAMP assay has the ability to specifically detect the virus in both diseased animals and infected epithelioma papilloma cyprinid (EPC) cells. The assay was shown to be 10-fold more sensitive than nested PCR and was able to detect concentrations of 10^鈭? (approximately 0.01 pg/渭L). The LAMP assay is relatively easy to perform in situ and the amplification products can be observed directly under UV light or via staining with SYBR Green I. The LAMP assay is also rapid and cost-effective. This study establishes the use of a LAMP assay for rapid detection of GSIV, which is a novel and important tool for the diagnosis of GSIV infection in laboratory or farmed Chinese giant salamanders.