Development and application of quantitative detection method for viral hemorrhagic septicemia virus (VHSV) genogroup IVa

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 nu...

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Veröffentlicht in:	Viruses 2014-05, Vol.6 (5), p.2204-2213
Hauptverfasser:	Kim, Jong-Oh, Kim, Wi-Sik, Kim, Si-Woo, Han, Hyun-Ja, Kim, Jin Woo, Park, Myoung Ae, Oh, Myung-Joo
Format:	Artikel
Sprache:	eng
Schlagworte:	Animals Cloning diagnostic Disease Efficiency Fish Fish Diseases - diagnosis Fish Diseases - virology Flatfishes - virology Genes Genomes Genotype Genotype & phenotype Infections Korea Mortality Novirhabdovirus - classification Novirhabdovirus - genetics Novirhabdovirus - isolation & purification Nucleocapsid - genetics olive flounder Paralichthys olivaceus Phylogenetics Pleuronectiformes Proteins quantitative detection real-time PCR Real-Time Polymerase Chain Reaction - methods Rhabdoviridae Infections - diagnosis Rhabdoviridae Infections - veterinary Rhabdoviridae Infections - virology Sepsis Time Factors Viral hemorrhagic septicemia virus viral hemorrhagic septicemia virus (VHSV) Viral Load - methods Viruses
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Zusammenfassung:	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.
ISSN:	1999-4915 1999-4915
DOI:	10.3390/v6052204