Sokolova T M, Selivanova T K, Lebedev A Iu, Bystrov N S, Gromashevskiĭ V L, Parasiuk N A, Ionova K S, Uryvaev L V
Vopr Virusol. 1996 Sep-Oct;41(5):209-14.
Genetic relationships of geographical isolates of the members of WEE virus serocomplex (McMillan, Fort Morgan, Highlands J, and Y62-33) were assessed by the polymerase chain reaction (PCR) and restriction analysis of the PCR products. Oligonucleotide primers (21 nucleotides in length) were chosen for NSP2, nucleocapsid C, and E2-E1 protein genes based on the known primary structure of the McMillan 16310-5614 genome (L. Uryvayev et al., 1994, 1995). These primers were shown to differentiate well the WEE and SV-like strains of the serocomplex. Y62-33 virus (Udmurtia, Russia) was identical to McMillan strain in three studied regions of NSP2, C, and E2-E1 genes. NSP2 gene could be detected in all the studied geographical isolates and was characterized by the same restriction patterns as endonucleases; it appeared to be the most conservative. The structural genes were less conservative. Fort Morgan virus (Colorado, USA) genome reliably differed from McMillan virus (California, USA) and was negative in PCR with primers to C and E2 gene regions. Highlands J genome (Florida, USA) was positive in PCR with the primers to E2-E1 gene regions but differed from McMillan strain by the nucleocapsid gene. An additional comparative PCR analysis of the C-E2 region in the McMillan and Highlands J genomes showed some, but not complete identity. The origin of these two viruses might be due to the selection of different forms of recombinant viruses. A good correlation of structural genes in PCR and the infectivity neutralization test was noted with the primers and polyclonal antibodies to the closely related strains. High specificity of PCR permits a more accurate detection of the virus origin and relationships.
通过聚合酶链反应(PCR)以及对PCR产物的限制性分析,评估了西部马脑炎病毒血清复合群成员(麦克米伦株、摩根堡株、高地J株和Y62 - 33株)地理分离株之间的遗传关系。基于麦克米伦16310 - 5614基因组已知的一级结构(L. 尤里瓦耶夫等人,1994年、1995年),针对非结构蛋白2(NSP2)、核衣壳蛋白C以及E2 - E1蛋白基因选择了长度为21个核苷酸的寡核苷酸引物。这些引物能够很好地区分血清复合群中的西部马脑炎病毒株和辛德毕斯病毒样株。Y62 - 33病毒(俄罗斯乌德穆尔特共和国)在NSP2、C以及E2 - E1基因的三个研究区域与麦克米伦株相同。在所有研究的地理分离株中都能检测到NSP2基因,并且其内切酶限制性图谱特征相同;它似乎是最保守的。结构基因的保守性较差。摩根堡病毒(美国科罗拉多州)的基因组与麦克米伦病毒(美国加利福尼亚州)有可靠的差异,并且用针对C基因区域和E2基因区域的引物进行PCR检测时呈阴性。高地J株基因组在用针对E2 - E1基因区域的引物进行PCR检测时呈阳性,但在核衣壳基因方面与麦克米伦株不同。对麦克米伦株和高地J株基因组中C - E2区域进行的额外比较PCR分析显示出一定程度但并非完全相同。这两种病毒的起源可能是由于选择了不同形式的重组病毒。在用针对密切相关毒株的引物和多克隆抗体进行PCR时,注意到结构基因与感染性中和试验之间有良好的相关性。PCR的高特异性使得能够更准确地检测病毒的起源和关系。