Kibenge F S, Lyaku J R, Rainnie D, Hammell K L
Department of Pathology and Microbiology, AVC Inc. and Department of Health Management(3), Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PEI, Canada.
J Gen Virol. 2000 Jan;81(Pt 1):143-50. doi: 10.1099/0022-1317-81-1-143.
Infectious salmon anaemia virus (ISAV) is a new orthomyxovirus-like virus. Thirteen isolates of ISAV (11 from Canada, one from Norway and one from Scotland) were studied for their replication in the CHSE-214 cell line compared with that in the SHK-1 cell line. All isolates replicated in SHK-1 cells, producing CPE between 3 and 12 days post-inoculation (p.i.). Six Canadian isolates also replicated in CHSE-214 cells, with production of CPE between 4 and 17 days p.i. Analysis of a one-step growth curve of ISAV in CHSE-214 cells showed that progeny virions remained predominantly cell-associated, accounting for the focalized nature of the CPE in the cell monolayer. One isolate (HKS 36) replicated in CHSE-214 cells, as shown by positive RT-PCR results of blind passages, but was non-cytopathic. All of the isolates were analysed for genetic heterogeneity by RT-PCR and RFLP with EcoRI and XhoI in a fraction of genome segment 2. The Canadian isolates showed a different RFLP profile to those of isolates Glesvaer/2/90 from Norway and 390/98 from Scotland. Structural proteins of four isolates, 'Back Bay 98', RPC/NB-877, RPC/NB-049 and Glesvaer/2/90, were examined further by SDS-PAGE. All viruses showed four major polypeptides, designated here as VP1-VP4, in Coomassie blue-stained gels. In isolates Glesvaer/2/90 and RPC/NB-877, these viral proteins had estimated molecular masses of 74, 53, 46 and 26.5 kDa, respectively. Viral proteins in isolates 'Back Bay 98' and RPC/NB-049 were of similar sizes, except that VP3 was 43 kDa. Taken together, these results show that there are phenotypic differences among strains of ISAV.
传染性鲑鱼贫血病毒（ISAV）是一种新的类正黏病毒。研究了13株ISAV（11株来自加拿大，1株来自挪威，1株来自苏格兰）在CHSE - 214细胞系中的复制情况，并与SHK - 1细胞系进行比较。所有分离株在SHK - 1细胞中均能复制，接种后3至12天出现细胞病变效应（CPE）。6株加拿大分离株在CHSE - 214细胞中也能复制，接种后4至17天出现CPE。对ISAV在CHSE - 214细胞中的一步生长曲线分析表明，子代病毒粒子主要与细胞相关，这解释了细胞单层中CPE的局灶性特征。一株分离株（HKS 36）在CHSE - 214细胞中能复制，盲传的RT - PCR结果呈阳性，但无细胞病变。通过RT - PCR以及用EcoRI和XhoI对基因组片段2的一部分进行限制性片段长度多态性（RFLP）分析，对所有分离株的遗传异质性进行了检测。加拿大分离株的RFLP图谱与来自挪威的Glesvaer/2/90分离株和来自苏格兰的390/98分离株不同。通过SDS - PAGE进一步检测了4株分离株“Back Bay 98”、RPC/NB - 877、RPC/NB - 049和Glesvaer/2/90的结构蛋白。在考马斯亮蓝染色的凝胶中，所有病毒均显示出4种主要多肽，在此命名为VP1 - VP4。在Glesvaer/2/90和RPC/NB - 877分离株中，这些病毒蛋白的估计分子量分别为74、53、46和26.5 kDa。“Back Bay 98”和RPC/NB - 049分离株中的病毒蛋白大小相似，只是VP3为43 kDa。综上所述，这些结果表明ISAV毒株之间存在表型差异。
