title: Development of a loop-mediated isothermal amplification assay for rapid detection of iridovirus in the Chinese giant salamander.
abstract: 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(-9) (approximately 0.01 pg/μL). 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.
authors.affiliation: Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, Hubei 430223, China; College of Fisheries, Huazhong Agricultural University, Wuhan 430071, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Meng
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Hongwei
authors.initials: H
authors.lastname: Liang
authors.forename: Lingbing
authors.initials: L
authors.lastname: Zeng
authors.forename: Hanbing
authors.initials: H
authors.lastname: Xiao
authors.forename: Congxin
authors.initials: C
authors.lastname: Xie
mesh_terms: D000818:Animals; D036022:Capsid Proteins / Q000235:genetics; D003365:Costs and Cost Analysis; D017931:DNA Primers / Q000235:genetics; D004266:DNA Virus Infections / Q000175:diagnosis / Q000662:veterinary*; D017994:Iridovirus / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014562:Caudata / Q000821:virology*; D014730:Veterinary Medicine / Q000191:economics / Q000379:methods*; D014773:Virology / Q000191:economics / Q000379:methods*
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers
keywords: Chinese giant salamander (Andrias davidianus); Iridovirus; Loop-mediated isothermal amplification (LAMP); Molecular detection