Viruses 2014, 6(5), 2204-2213; https://doi.org/10.3390/v6052204
Development and Application of Quantitative Detection Method for Viral Hemorrhagic Septicemia Virus (VHSV) Genogroup IVa
1
Department of Aqualife Medicine, Chonnam National University, Yeosu 550-749, Korea
2
Aquatic Life Disease Control Division, National Fisheries Research and Development Institute, Busan 619-902, Korea
*
Author to whom correspondence should be addressed.
Received: 7 April 2014 / Revised: 8 May 2014 / Accepted: 12 May 2014 / Published: 23 May 2014
Abstract
Viral hemorrhagic septicemia virus (VHSV) is a problematic pathogen in olive flounder (Paralichthys olivaceus) aquaculture farms in Korea. Thus, it is necessary to develop a rapid and accurate diagnostic method to detect this virus. We developed a quantitative RT-PCR (qRT-PCR) method based on the nucleocapsid (N) gene sequence of Korean VHSV isolate (Genogroup IVa). The slope and R2 values of the primer set developed in this study were −0.2928 (96% efficiency) and 0.9979, respectively. Its comparison with viral infectivity calculated by traditional quantifying method (TCID50) showed a similar pattern of kinetic changes in vitro and in vivo. The qRT-PCR method reduced detection time compared to that of TCID50, making it a very useful tool for VHSV diagnosis. View Full-TextKeywords:
viral hemorrhagic septicemia virus (VHSV); quantitative detection; real-time PCR; diagnostic; olive flounder
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