Abstract：
White sturgeon Acipenser transmontanus is the primary species used for caviar and sturgeon meat production in the USA. An important pathogen of white sturgeon is acipenserid herpesvirus 2 (AciHV-2). In this study, 4 archived isolates from temporally discrete natural outbreaks spanning the past 30 yr were sequenced via Illumina and Oxford Nanopore Technologies platforms. Assemblies of approximately 134 kb were obtained for each isolate, and the putative ATPase subunit of the terminase gene was selected as a potential quantitative PCR (qPCR) target based on sequence conservation among AciHV-2 isolates and low sequence homology with other important viral pathogens. The qPCR was repeatable and reproducible, with a linear dynamic range covering 5 orders of magnitude, an efficiency of approximately 96%, an R2 of 0.9872, and an analytical sensitivity of 103 copies per reaction after 35 cycles. There was no cross-reaction with other known viruses or closely related sturgeon species, and no inhibition by sturgeon DNA. Clinical accuracy was assessed from white sturgeon juveniles exposed to AciHV-2 by immersion. Viral culture (gold standard) and qPCR were in complete agreement for both cell culture negative and cell culture positive samples, indicating that this assay has 100% relative accuracy compared to cell culture during an active outbreak. The availability of a whole-genome sequence for AciHV-2 and a highly specific and sensitive qPCR assay for detection of AciHV-2 in white sturgeon lays a foundation for further studies on host-pathogen interactions while providing a specific and rapid test for AciHV-2 in captive and wild populations.
摘要：
白色st鱼Acipensertransmontanus是美国用于鱼子酱和st鱼肉生产的主要物种。白色st鱼的重要病原体是阿西丝样疱疹病毒2型（AciHV-2）。在这项研究中,通过Illumina和OxfordNanoporeTechnologies平台对过去30年中时间离散的自然爆发的4个存档分离物进行了测序。每个分离株获得约134kb的组件，并且基于AciHV-2分离株之间的序列保守性和与其他重要病毒病原体的低序列同源性，选择了末端酶基因的推定ATPase亚基作为潜在的定量PCR（qPCR）靶标。qPCR是可重复和可重复的，具有覆盖5个数量级的线性动态范围，效率约为96%，R2为0.9872，35个循环后每个反应的分析灵敏度为103个拷贝。与其他已知病毒或密切相关的st鱼物种没有交叉反应，并且没有被st鱼DNA抑制。通过浸入暴露于AciHV-2的白色st鱼幼鱼评估了临床准确性。对于细胞培养阴性和细胞培养阳性样品，病毒培养（金标准）和qPCR完全一致。表明该测定与活跃爆发期间的细胞培养相比具有100%的相对准确性。AciHV-2全基因组序列的可用性以及用于检测白st中AciHV-2的高度特异性和灵敏的qPCR检测方法的可用性为进一步研究宿主与病原体的相互作用奠定了基础，同时为圈养和野生种群中的AciHV-2提供了特异性和快速的测试。
