Alfieri A A, Leite J P, Nakagomi O, Kaga E, Woods P A, Glass R I, Gentsch J R
Laboratory of Animal Virology, Londrina State University, Brazil.
Arch Virol. 1996;141(12):2353-64. doi: 10.1007/BF01718636.
We report the molecular characterization of rotavirus genotype P[8]G5 strains found in fecal specimens collected in four different regions of Brazil, using digoxigenin(dig)-labeled oligonucleotide probes, sequence analysis, and RNA-RNA hybridization. The closest sequence relationships of the neutralization antigens of these strains were to the VP4 protein of P1A[8]G1 strain KU (93.3% identity in amino acids 11 to 282) and to the VP7 protein of G serotype 5 strain OSU (87.6% identity in amino acids 8 to 232). Based on VP7 sequence differences, we designed dig-probes that allowed us to discriminate porcine OSU-like strains from G5 strains isolated from Brazilian infants. The genetic relationships of two P[8]G5 isolates to other rotavirus genogroups were analyzed by RNA-RNA hybridization with [32P]-GTP probes representative of serotypes P1A[8]G1 (Wa), P[8]G3 (AU17), and P9[7]G5 (OSU). The Brazilian P[8]G5 strains showed sequence homology with genes of Wa-like and OSU-like strains, suggesting that these two strains were naturally occurring reassortants between members of the Wa and porcine rotavirus genogroups. The identification of these strains in diverse geographic areas of Brazil underscores their stability and demonstrates the emergence of clinically important rotavirus diarrhea strains by reassortment.
我们报告了在巴西四个不同地区收集的粪便标本中发现的轮状病毒基因型P[8]G5毒株的分子特征,采用了地高辛(dig)标记的寡核苷酸探针、序列分析和RNA-RNA杂交技术。这些毒株中和抗原的最接近序列关系与P1A[8]G1毒株KU的VP4蛋白(氨基酸11至282的同一性为93.3%)以及G血清型5毒株OSU的VP7蛋白(氨基酸8至232的同一性为87.6%)相关。基于VP7序列差异,我们设计了地高辛探针,使我们能够区分猪源OSU样毒株与从巴西婴儿中分离出的G5毒株。通过与代表血清型P1A[8]G1(Wa)、P[8]G3(AU17)和P9[7]G5(OSU)的[32P]-GTP探针进行RNA-RNA杂交,分析了两个P[8]G5分离株与其他轮状病毒基因组的遗传关系。巴西的P[8]G5毒株与Wa样和OSU样毒株的基因显示出序列同源性,表明这两个毒株是Wa和猪轮状病毒基因组成员之间自然发生的重配体。在巴西不同地理区域鉴定出这些毒株,突出了它们的稳定性,并证明了通过重配出现具有临床重要性的轮状病毒腹泻毒株。