Tattiyapong P, Sirikanchana K, Surachetpong W
Center for Advanced Studies for Agriculture and Food (CASAF), Kasetsart University Institute for Advanced Studies, Kasetsart University (NRU-KU), Bangkok, Thailand.
Department of Veterinary Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok, Thailand.
J Fish Dis. 2018 Feb;41(2):255-261. doi: 10.1111/jfd.12708. Epub 2017 Oct 13.
Tilapia lake virus (TiLV) 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 TiLV in clinical samples and experimentally challenged fish. All 30 field samples with clinical signs and history consistent with TiLV infection were positive for TiLV 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 TiLV 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 TiLV viruses that facilitates active surveillance programme and disease containment.
罗非鱼湖病毒（TiLV）是一种新出现的病原体，与不同国家野生和养殖罗非鱼的高死亡率有关。在本研究中，开发了一种基于SYBR Green的逆转录定量聚合酶链反应（RT-qPCR）检测方法，用于靶向该病毒的第三片段，以检测和定量临床样本及实验感染鱼中的TiLV。通过所开发的RT-qPCR方法检测，所有30份具有与TiLV感染相符的临床症状和病史的现场样本均为TiLV阳性。RT-qPCR技术在病毒检测方面分别比RT-PCR和细胞培养法中的病毒分离法敏感100倍和10000倍。RT-qPCR方法的检测限低至2个病毒拷贝/μl。此外，RT-qPCR技术可用于检测包括鳃、肝、脑、心脏、前肾和脾脏在内的各种鱼类组织中的TiLV。重要的是，本研究提供了一种准确可靠的快速检测TiLV病毒的方法，有助于开展主动监测计划和疾病防控。
