Midthun K, Flores J, Taniguchi K, Urasawa S, Kapikian A Z, Chanock R M
J Clin Microbiol. 1987 Jul;25(7):1269-74. doi: 10.1128/jcm.25.7.1269-1274.1987.
Antigenic characterization of human rotaviruses by plaque reduction neutralization assay has revealed four distinct serotypes. The outer capsid protein VP7, coded for by gene 8 or 9, is a major neutralization protein; however, studies of rotaviruses derived from genetic reassortment between two strains have confirmed that another outer capsid protein, VP3, is in some cases equally important in neutralization. In this study, the genetic relatedness of the genes coding for VP7 of human rotaviruses belonging to serotypes 1 through 4 was examined by hybridization of their denatured double-stranded genomic RNAs to labeled single-stranded mRNA probes derived from human-animal rotavirus reassortants containing only the VP7 gene of their human rotavirus parent. A high degree of homology was demonstrated between the VP7 genes of strain D and other serotype 1 human rotaviruses, strain DS-1 and other serotype 2 human rotaviruses, strain P and other serotype 3 human rotaviruses, and strain ST3 and other serotype 4 human rotaviruses. Hybrid bands could not be demonstrated between the VP7 gene of D, DS-1, P, or ST3 and the corresponding gene of human rotaviruses belonging to a different serotype. RNA specimens extracted from the stools of 15 Venezuelan children hospitalized with rotavirus diarrhea were hybridized to each of the reassortant probes representing the four human serotypes. All five viruses with short RNA patterns showed homology with the DS-1 strain VP7 gene; two of these were previously adapted to tissue culture and shown to be serotype 2 strains by tissue culture neutralization. Of the remaining 10 viruses with long RNA patterns, 2 hybridized only to the D strain VP7 gene, 6 hybridized only to the P strain VP7 gene, and 2 hybridized only to the ST3 strain VP7 gene. Hybridization using single human rotavirus gene substitution reassortants as probes may provide an alternative method for identifying the VP7 serotype of field isolates that would circumvent the need for tissue culture adaptation.
通过蚀斑减少中和试验对人轮状病毒进行抗原特性分析，已揭示出四种不同的血清型。由基因8或9编码的外衣壳蛋白VP7是主要的中和蛋白；然而，对来自两株病毒之间基因重配的轮状病毒的研究证实，另一种外衣壳蛋白VP3在某些情况下对中和作用同样重要。在本研究中，通过将人轮状病毒1至4型的变性双链基因组RNA与源自仅含其人轮状病毒亲本VP7基因的人-动物轮状病毒重配体的标记单链mRNA探针杂交，检测了编码VP7的基因的遗传相关性。结果表明，D株与其他1型人轮状病毒、DS-1株与其他2型人轮状病毒、P株与其他3型人轮状病毒、ST3株与其他4型人轮状病毒的VP7基因之间具有高度同源性。在D株、DS-1株、P株或ST3株的VP7基因与属于不同血清型的人轮状病毒的相应基因之间未显示杂交带。从15名因轮状病毒腹泻住院的委内瑞拉儿童粪便中提取的RNA标本，与代表四种人血清型的每种重配探针进行杂交。所有五种具有短RNA模式的病毒均与DS-1株VP7基因显示同源性；其中两株先前已适应组织培养，并通过组织培养中和试验显示为2型毒株。在其余10种具有长RNA模式的病毒中，2株仅与D株VP7基因杂交，6株仅与P株VP7基因杂交，2株仅与ST3株VP7基因杂交。使用单个人轮状病毒基因替代重配体作为探针进行杂交，可能为鉴定现场分离株的VP7血清型提供一种替代方法，从而无需进行组织培养适应。
