Cornwell Emily R, Eckerlin Geofrey E, Getchell Rodman G, Groocock Geoffrey H, Thompson Tarin M, Batts William N, Casey Rufina N, Kurath Gael, Winton James R, Bowser Paul R, Bain Mark B, Casey James W
Aquatic Animal Health Program, Department of Microbiology and Immunology, College of Veterinary Medicine, Upper Tower Road, Cornell University, Ithaca, New York 14853, USA.
J Aquat Anim Health. 2011 Dec;23(4):207-17. doi: 10.1080/08997659.2011.644411.
Viral hemorrhagic septicemia virus (VHSV) was first detected in the Laurentian Great Lakes in 2005 during a mortality event in the Bay of Quinte, Lake Ontario. Subsequent analysis of archived samples determined that the first known isolation of VHSV in the Laurentian Great Lakes was from a muskellunge Esox masquinongy collected in Lake St. Clair in 2003. By the end of 2008, mortality events and viral isolations had occurred in all of the Laurentian Great Lakes except Lake Superior. In 2009, a focused disease surveillance program was designed to determine whether VHSV was also present in Lake Superior. In this survey, 874 fish from 7 sites along the U.S. shoreline of Lake Superior were collected during June 2009. Collections were focused on nearshore species known to be susceptible to VHSV. All fish were dissected individually by using aseptic techniques and were tested for the presence of VHSV genetic material by use of a quantitative reverse transcription (qRT) polymerase chain reaction (PCR) targeting the viral nucleoprotein gene. Seventeen fish from two host species at two different sites tested positive at low levels for VHSV. All attempts to isolate virus in cell culture were unsuccessful. However, the presence of viral RNA was confirmed independently in five fish by using a nested PCR that targeted the glycoprotein (G) gene. Partial G gene sequences obtained from three fish were identical to the corresponding sequence from the original 2003 VHSV isolate (MI03) from muskellunge. These detections represent the earliest evidence for the presence of VHSV in Lake Superior and illustrate the utility of the highly sensitive qRT-PCR assay for disease surveillance in aquatic animals.
2005年，在安大略湖昆特湾发生的一次死亡事件中，首次在劳伦琴大湖检测到病毒性出血性败血症病毒（VHSV）。随后对存档样本的分析确定，劳伦琴大湖首次已知的VHSV分离株来自2003年在圣克莱尔湖采集的一条北美狗鱼。到2008年底，除苏必利尔湖外，所有劳伦琴大湖均发生了死亡事件并分离出了病毒。2009年，设计了一项重点疾病监测计划，以确定苏必利尔湖中是否也存在VHSV。在这项调查中，2009年6月从苏必利尔湖美国沿岸的7个地点采集了874条鱼。采集重点是已知易感染VHSV的近岸物种。所有鱼均采用无菌技术单独解剖，并使用针对病毒核蛋白基因的定量逆转录（qRT）聚合酶链反应（PCR）检测VHSV遗传物质的存在。来自两个不同地点的两个宿主物种的17条鱼对VHSV呈低水平阳性检测。在细胞培养中分离病毒的所有尝试均未成功。然而，通过使用针对糖蛋白（G）基因的巢式PCR，在五条鱼中独立确认了病毒RNA的存在。从三条鱼中获得的部分G基因序列与2003年从北美狗鱼分离出的原始VHSV分离株（MI03）的相应序列相同。这些检测结果代表了苏必利尔湖存在VHSV的最早证据，并说明了高灵敏度qRT-PCR检测法在水生动物疾病监测中的实用性。
