Santi Nina, Vakharia Vikram N, Evensen Øystein
Department of Pathology, National Veterinary Institute, Oslo, Norway.
Virology. 2004 Apr 25;322(1):31-40. doi: 10.1016/j.virol.2003.12.016.
Infectious pancreatic necrosis viruses (IPNVs) belonging to the family Birnaviridae display a high degree of antigenic variability, pathogenicity, and differences in outbreak mortality in salmonid species. To determine if virus isolates of Sp serotype differ in virulence, fry of Atlantic salmon (Salmo salar L.) were challenged with nine different field strains. These viruses caused either high mortality and severe pathological changes or low mortality and no lesions. To study the molecular basis for the variation in virulence of IPNV, complete nucleotide sequences of segment A of all these strains as well as segment B of three selected strains were determined. All viruses tested had a unique genome sequence. Only minor differences were noted in the genes encoding VP1, VP3, and VP4 proteins, whereas most changes were observed in the gene encoding the VP2 protein. A high level of variation was found in the small open reading frame (ORF), which encodes a 15-kDa nonstructural (NS) polypeptide also known as VP5. One of the strains lacked the initiation codon for this protein, whereas the other four could encode a truncated version of the NS protein. Additional data obtained by sequencing of the NS and VP2 genes directly from diseased fish demonstrated changes in the VP2 gene after two passages in cell culture, which could possibly be associated with attenuation. Comparison of the deduced amino acid sequences of the NS and VP2 genes reveals that the virulent strains possess a 12-kDa coding NS gene and have residues Thr, Ala, Thr/Ala, and Tyr/His at positions 217, 221, 247, and 500 of the VP2 gene, respectively-the motifs identified in this study to be involved in the virulence of IPNV.
属于双RNA病毒科的传染性胰腺坏死病毒(IPNV)在鲑科鱼类中表现出高度的抗原变异性、致病性以及暴发死亡率差异。为了确定Sp血清型的病毒分离株在毒力上是否存在差异,用九种不同的野外毒株对大西洋鲑(Salmo salar L.)鱼苗进行了攻毒试验。这些病毒要么导致高死亡率和严重的病理变化,要么导致低死亡率且无病变。为了研究IPNV毒力变异的分子基础,测定了所有这些毒株A片段以及三个选定毒株B片段的完整核苷酸序列。所有测试病毒都有独特的基因组序列。在编码VP1、VP3和VP4蛋白的基因中仅发现微小差异,而在编码VP2蛋白的基因中观察到的变化最多。在编码一种15 kDa非结构(NS)多肽(也称为VP5)的小开放阅读框(ORF)中发现了高度变异。其中一个毒株缺少该蛋白的起始密码子,而其他四个毒株可以编码截短版的NS蛋白。直接从患病鱼中对NS和VP2基因进行测序获得的其他数据表明,在细胞培养中传代两次后VP2基因发生了变化,这可能与病毒减毒有关。对NS和VP2基因推导的氨基酸序列进行比较发现,强毒株拥有一个编码12 kDa NS的基因,并且在VP2基因的第217、221、247和500位分别具有苏氨酸、丙氨酸、苏氨酸/丙氨酸和酪氨酸/组氨酸残基——本研究中确定的与IPNV毒力有关的基序。