Bootland L M, Leong J A
Laboratory for Fish Disease Research, Oregon State University, Corvallis 97331-3804.
Appl Environ Microbiol. 1992 Jan;58(1):6-13. doi: 10.1128/aem.58.1.6-13.1992.
A staphylococcal coagglutination test was developed for the rapid detection of infectious hematopoietic necrosis virus (IHNV) in cell cultures and infected fish. The test could be completed in 15 min but required a minimum IHNV titer of 10(6) PFU/ml to obtain a positive reaction. All IHNV isolates, representing the five electropherotypes taken from a wide variety of species and different geographic ranges, caused coagglutination of Staphylococcus aureus cells sensitized with rabbit polyclonal serum against the Round Butte IHNV isolate. The coagglutination reaction was blocked by preincubation of IHNV with homologous antiserum, and IHNV did not cause coagglutination of S. aureus cells sensitized with normal rabbit serum. In specificity tests, cells sensitized with rabbit anti-IHNV serum or normal serum did not coagglutinate in the presence of infectious pancreatic necrosis virus, viral hemorrhagic septicemia virus, cell culture medium components, or media from cultures of cell lines of salmonid and nonsalmonid origin. Most importantly, the coagglutination test was able to detect and identify IHNV directly from experimentally infected rainbow trout fry, the organs of naturally infected adult kokanee salmon and winter steelhead trout, and ovarian fluids of the winter steelhead trout. The coagglutination test is very suitable for field use, since it is inexpensive, simple to interpret, sensitive, and rapid and requires no specialized equipment.
已开发出一种葡萄球菌协同凝集试验，用于快速检测细胞培养物和受感染鱼类中的传染性造血坏死病毒（IHNV）。该试验可在15分钟内完成，但要获得阳性反应，所需的IHNV最低滴度为10(6) PFU/ml。所有代表从多种物种和不同地理区域获取的五种电泳型的IHNV分离株，均能使经兔抗Round Butte IHNV分离株多克隆血清致敏的金黄色葡萄球菌细胞发生协同凝集。IHNV与同源抗血清预孵育可阻断协同凝集反应，且IHNV不会使经正常兔血清致敏的金黄色葡萄球菌细胞发生协同凝集。在特异性试验中，在传染性胰腺坏死病毒、病毒性出血性败血症病毒、细胞培养基成分或鲑科和非鲑科来源细胞系培养物的培养基存在的情况下，经兔抗IHNV血清或正常血清致敏的细胞不会发生协同凝集。最重要的是，协同凝集试验能够直接从实验感染的虹鳟鱼苗、自然感染的成年红大马哈鱼和冬虹鳟的器官以及冬虹鳟的卵巢液中检测和鉴定IHNV。该协同凝集试验非常适合现场使用，因为它价格低廉、易于解读、灵敏、快速，且无需专门设备。
