Department of Comparative Biosciences, College of Veterinary Medicine, University of Illinois, 2001 South Lincoln Avenue, Urbana, IL 61802, USA.
J Virol Methods. 2013 Mar;188(1-2):121-5. doi: 10.1016/j.jviromet.2012.12.012. Epub 2012 Dec 26.
Ranavirus has caused disease epidemics and mass mortality events globally in free-ranging fish, amphibian, and reptile populations. Viral isolation and conventional PCR are the most common methods for diagnosis. In this study, a quantitative real-time PCR (qPCR) assay was developed using a TaqMan probe-based assay targeting a highly conserved region of the major capsid protein of frog virus 3-like virus (FV3-like) (Family Iridoviridae, genera Ranavirus). Standard curves were generated from a viral DNA segment cloned within a plasmid. The assay detected viral DNA 1000 times lower than conventional PCR. Thirty-one clinical samples (whole blood and oral swabs) from box turtles were tested using these assays and the prevalence of the virus determined. Quantitative PCR allows for a superior, rapid, sensitive, and quantitative method for detecting FV3-like virus in box turtles, and this assay will be useful for early detection and disease monitoring.
蛙病毒 3 样病毒（类虹彩病毒科，蛙病毒属）的主要衣壳蛋白的高度保守区为靶标，建立了一种基于 TaqMan 探针的定量实时 PCR（qPCR）检测方法。该方法采用克隆于质粒的病毒 DNA 片段制作标准曲线。该检测方法的检测下限比常规 PCR 低 1000 倍。用该方法和检测对 31 份箱龟的临床样本（全血和口腔拭子）进行了检测，并确定了病毒的流行率。qPCR 可用于箱龟中 FV3 样病毒的快速、灵敏、定量检测，该方法将有助于早期检测和疾病监测。
