Premium
Development and validation of a reverse transcription quantitative polymerase chain reaction for tilapia lake virus detection in clinical samples and experimentally challenged fish
Author(s) -
Tattiyapong P,
Sirikanchana K,
Surachetpong W
Publication year - 2018
Publication title -
journal of fish diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.819
H-Index - 85
eISSN - 1365-2761
pISSN - 0140-7775
DOI - 10.1111/jfd.12708
Subject(s) - biology , reverse transcription polymerase chain reaction , reverse transcriptase , virus , tilapia , gill , virology , pathogen , real time polymerase chain reaction , polymerase chain reaction , spleen , detection limit , specific pathogen free , microbiology and biotechnology , fish <actinopterygii> , messenger rna , gene , fishery , chromatography , immunology , chemistry , biochemistry
Tilapia lake virus (Ti LV ) is an emerging pathogen associated with high mortalities of wild and farm‐raised tilapia in different countries. In this study, a SYBR green‐based reverse transcription quantitative polymerase chain reaction ( RT ‐ qPCR ) assay targeting segment three of the virus was developed to detect and quantify Ti LV in clinical samples and experimentally challenged fish. All 30 field samples with clinical signs and history consistent with Ti LV infection were positive for Ti LV as detected by the developed RT ‐ qPCR method. The RT ‐ qPCR technique provided 100 and 10,000 times more sensitive for virus detection than those offered by the RT ‐ PCR and virus isolation in cell culture methods, respectively. The detection limit of the RT ‐ qPCR method was as low as two viral copies/μl. Moreover, the RT ‐ qPCR technique could be applied for Ti LV detection in various fish tissues including gills, liver, brain, heart, anterior kidney and spleen. Significantly, this study delivered an accurate and reliable method for rapid detection of Ti LV viruses that facilitates active surveillance programme and disease containment.