Teige Lena Hammerlund, Lund Morten, Haatveit Hanne M, Røsæg Magnus Vikan, Wessel Øystein, Dahle Maria K, Storset Anne K
Department of Food Safety and Infection Biology, Norwegian University of Life Sciences, N-0454 Oslo, Norway.
Norwegian Veterinary Institute, N-0454 Oslo, Norway.
Fish Shellfish Immunol. 2017 Apr;63:491-499. doi: 10.1016/j.fsi.2017.02.043. Epub 2017 Feb 27.
Future growth in aquaculture relies strongly on the control of diseases and pathogens. Vaccination has been a successful strategy for obtaining control of bacterial diseases in fish, but for viral diseases, vaccine development has been more challenging. Effective long-term protection against viral infections is not yet fully understood for fish, and in addition, optimal tools to monitor adaptive immunity are limited. Assays that can detect specific antibodies produced in response to viral infection in fish are still in their early development. Multiplex bead based assays have many advantages over traditional assays, since they are more sensitive and allow detection of multiple antigen-specific antibodies simultaneously in very small amounts of plasma or serum. In the present study, a bead based assay have been developed for detection of plasma IgM directed against Piscine orthoreovirus (PRV), the virus associated with the disease Heart and skeletal muscle inflammation (HSMI) in farmed Atlantic salmon. Using recombinant PRV proteins coated on beads, antibodies targeting the structural outer capsid protein μ1 and the non-structural protein μNS were detected. Results from a PRV cohabitation challenge trial indicated that the antibody production was initiated approximately two weeks after the peak phase of PRV infection, coinciding with typical HSMI pathology. Thereafter, the antibody production increased while the epicardial inflammation became less prominent. In conclusion, the novel assay can detect PRV-specific antibodies that may play a role in viral defence. The bead-based immunoassay represents a valuable tool for studies on HSMI and possibly other diseases in aquaculture.
水产养殖的未来增长在很大程度上依赖于疾病和病原体的控制。疫苗接种一直是控制鱼类细菌性疾病的成功策略，但对于病毒性疾病，疫苗开发更具挑战性。鱼类针对病毒感染的有效长期保护机制尚未完全明确，此外，监测适应性免疫的最佳工具也很有限。能够检测鱼类因病毒感染而产生的特异性抗体的检测方法仍处于早期发展阶段。基于多重微珠的检测方法比传统检测方法有许多优势，因为它们更灵敏，能够在极少量的血浆或血清中同时检测多种抗原特异性抗体。在本研究中，开发了一种基于微珠的检测方法，用于检测针对鱼类正呼肠孤病毒（PRV）的血浆IgM，该病毒与养殖大西洋鲑鱼的心脏和骨骼肌炎症（HSMI）疾病有关。使用包被在微珠上的重组PRV蛋白，检测到了靶向结构外衣壳蛋白μ1和非结构蛋白μNS的抗体。PRV同居攻毒试验的结果表明，抗体产生在PRV感染高峰期后约两周开始，这与典型的HSMI病理变化一致。此后，抗体产生增加，而心外膜炎症变得不那么明显。总之，这种新型检测方法可以检测可能在病毒防御中起作用的PRV特异性抗体。基于微珠的免疫分析是研究HSMI以及水产养殖中其他可能疾病的有价值工具。
