Crockford Melanie, Jones John B, Crane Mark S J, Wilcox Graham E
Department of Fisheries Western Australia, PO Box 20, North Beach, Western Australia 6920, Australia.
Dis Aquat Organ. 2005 Dec 30;68(1):1-5. doi: 10.3354/dao068001.
In 1995, and again in 1998, high mortalities of pilchards Sardinops sagax neopilchardus were seen over their entire geographical range on the Australian coastline. A virus with typical herpesvirus morphology was identified as the causative agent, although the source of the virus remains unknown. At the time of the mortality events, the only available diagnostic test available for the detection of the virus was electron microscopy and hence, development of a rapid diagnostic test for detection and identification of the virus was required. Initial sequence data for Pilchard herpesvirus (PHV) was acquired by comparing the highly conserved region of the ORF 62 gene of other piscine herpesviruses (Ictalurid herpesvirus 1 [Channel catfish virus, CCV] and Salmonid herpesvirus 2 [Oncorhynchus masou virus, SaHV2, OMV]), and designing primers that successfully amplified a fragment of PHV. Here we describe the use of polymerase chain reaction (PCR) technology to detect PHV in infected tissues. Sequence analysis of amplified fragment resulting from this PCR is different from all known herpesviruses and can therefore distinguish PHV from all known viruses.
1995年以及1998年,在澳大利亚海岸线的整个地理范围内,均出现了新pilchard(Sardinops sagax neopilchardus)的高死亡率情况。一种具有典型疱疹病毒形态的病毒被确定为病原体,尽管该病毒的来源仍然未知。在死亡率事件发生时,唯一可用于检测该病毒的诊断测试是电子显微镜检查,因此,需要开发一种用于检测和鉴定该病毒的快速诊断测试。通过比较其他鱼类疱疹病毒(斑点叉尾鮰疱疹病毒1[通道鲶鱼病毒,CCV]和鲑鱼疱疹病毒2[马苏大麻哈鱼病毒,SaHV2,OMV])的ORF 62基因的高度保守区域,获取了pilchard疱疹病毒(PHV)的初始序列数据,并设计出成功扩增PHV片段的引物。在此,我们描述了使用聚合酶链反应(PCR)技术在受感染组织中检测PHV的方法。该PCR扩增片段的序列分析不同于所有已知疱疹病毒,因此可以将PHV与所有已知病毒区分开来。