Soliman H, El-Matbouli M
Clinical Division of Fish Medicine, University of Veterinary Medicine, Vienna, Austria.
J Fish Dis. 2018 May;41(5):761-772. doi: 10.1111/jfd.12774. Epub 2018 Jan 9.
Carp oedema virus (CEV) and koi herpes virus (KHV) are of major concern to common carp breeders and koi enthusiasts worldwide. The viruses cause diseases that exhibit similar external signs; thus, it is difficult to distinguish between them clinically. In this study, we developed and optimized rapid and accurate single- and multiplex isothermal diagnostic tools, based on recombinase polymerase amplification (RPA), for detection and differentiation of CEV and KHV. The assays were combined with a lateral flow dipstick to enable visual detection of amplification products and simplify post-amplification analysis. Both CEV- and KHV-RPA assays were specific for their target virus. The lower detection limits of the assays were similar to those of established diagnostic PCR tests for the viruses. A sample preparation method was optimized to eliminate the need for total DNA extraction from fish tissues. The estimated time to perform these RPA assays, from receiving the sample to having a result, is 50 min, compared to 10 and 7 hr for CEV- and KHV-PCR tests, respectively. The assays can be performed in field situations to improve screening of fish and reduce spread of these viruses and thereby enhance the common carp and koi industries.
鲤鱼水肿病毒（CEV）和锦鲤疱疹病毒（KHV）是全球鲤鱼养殖者和锦鲤爱好者主要关注的对象。这些病毒引发的疾病具有相似的外部症状；因此，临床上很难区分它们。在本研究中，我们基于重组酶聚合酶扩增技术（RPA）开发并优化了快速且准确的单重和多重等温诊断工具，用于检测和区分CEV与KHV。这些检测方法与侧向流动试纸条相结合，以便能够直观地检测扩增产物并简化扩增后分析。CEV和KHV的RPA检测方法对各自的目标病毒均具有特异性。这些检测方法的最低检测限与针对这些病毒的既定诊断性PCR检测方法相似。优化了一种样品制备方法，无需从鱼组织中提取总DNA。进行这些RPA检测从接收样品到得出结果的估计时间为50分钟，相比之下，CEV和KHV的PCR检测分别需要10小时和7小时。这些检测方法可在现场进行，以改进对鱼类的筛查并减少这些病毒的传播，从而促进鲤鱼和锦鲤产业的发展。
