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http://dx.doi.org/10.7847/jfp.2012.25.3.257

Evaluation of reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay for detection of infectious hematopoietic necrosis virus (IHNV)  

Kim, Wi-Sik (The Fisheries Science Institute, Chonnam National University)
Jeon, Chan-Hyeok (Department of Marine Bioscience, Gangneung-Wonju National University)
Kim, Jeong-Ho (Department of Marine Bioscience, Gangneung-Wonju National University)
Oh, Myung-Joo (Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University)
Publication Information
Journal of fish pathology / v.25, no.3, 2012 , pp. 257-262 More about this Journal
Abstract
A reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay was evaluated to monitor infectious hematopoietic necrosis virus (IHNV) from artificially infected rainbow trout Oncorhynchus mykiss. The cumulative mortalities of fish challenged with IHNV at $10^{6.5}\;TCID_{50}$/fish, $10^{5.5}\;TCID_{50}$/fish and $10^{4.5}\;TCID_{50}$/fish were 40%, 0% and 0%, respectively. Dead fish and survivors at 16 and 28 d post-challenge in each group were employed for IHNV detection by RT-LAMP assay and virus isolation using BF-2 cells. IHNV from $10^{4.3}$ to $10^{6.8}\;TCID_{50}/ml$ was isolated from all the dead fish and also detected in all of the examined dead fish by RT-LAMP assay. In survivors at 16 d, 60% (3/5 fish, $10^{2.8}-10^{5.05}\;TCID_{50}/ml$), 20% (1/5 fish, $10^{1.05}\;TCID_{50}/ml$) and 60% (3/5 fish, $10^{1.05}-10^{4.8}\;TCID_{50}/ml$) were found to be IHNV-positive by virus isolation in fish challenged with IHNV at $10^{6.5}\;TCID_{50}$/fish, $10^{5.5}\;TCID_{50}$/fish and $10^{4.5}\;TCID_{50}$/fish, respectively, while 20% (1/5 fish), 0% (0/5 fish) and 20% (1/5 fish) were IHNV-positive by RT-LAMP assay. No IHNV was detected in the survivors at 28 d and control fish. These results indicate that the RT-LAMP assay is useful for detection of IHNV in diseased fish although it is not enough to monitor virus in IHNV-survivors.
Keywords
IHNV; Rainbow trout; RT-LAMP;
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