Lu W, Duhamel G E, Benfield D A, Grotelueschen D M
Department of Veterinary and Biomedical Sciences, University of Nebraska-Lincoln 68583-0905.
Vet Microbiol. 1994 Nov;42(2-3):159-70. doi: 10.1016/0378-1135(94)90015-9.
Two strains of bovine rotavirus (BRV), designated strain Nebraska Scottsbluff-1 (NS-1) and NS-2, were isolated from 2 neighboring cow-calf beef cattle ranches where dams had been vaccinated with a commercial vaccine containing group A BRV strain Neonatal Calf Diarrhea Virus (NCDV)-Lincoln (P1:G6). Northern blot hybridizations using whole genomic RNA probes indicated that strains NS-1 and NS-2 had identical group A RNA electrophoretic patterns and were homologous at all gene segments. Strain NS-1 was compared with reference group A BRV strains using serological and genotypic methods. In vitro virus neutralization assays indicated that strain NS-1 was neutralized by a G6-specific neutralizing monoclonal antibody (mAb) and guinea pig hyperimmune serum (GPHS) raised against BRV strain B641 (P5:G6), but not by G10-specific neutralizing mAb or GPHS raised against BRV strain B223 (P11:G10). Nucleic acid hybridization experiments using whole-genomic RNA probes revealed that gene segment 4 of strain NS-1 differed from BRV strains NCDV-Lincoln and B223, but hybridized with strain B641. Conversely, gene segment 5 of strain NS-1 hybridized with BRV strain B223, but not with BRV strains NCDV-Lincoln and B641. A G-specific cDNA probe produced by reverse transcription polymerase chain reaction (RT-PCR) amplification of strain NS-1 hybridized specifically only with G6 strains NCDV-Lincoln and B641, but not with G10 strain B223. Co-electrophoresis experiments using strains NS-1, B641, and B223 further confirmed these results, suggesting that strain NS-1 was a naturally-occurring reassortant BRV between strains B641 and B223. Taken together these results indicated that a naturally-occurring group A BRV reassortant with a P gene different from the vaccine virus was responsible for the diarrheal syndrome observed on both ranches. Results from this study also indicate the existence of at least 2 different gene segments 5 among group A BRV infecting cattle.
从两个相邻的肉牛饲养场分离出两株牛轮状病毒（BRV），分别命名为内布拉斯加斯科茨布拉夫 -1 株（NS-1）和 NS-2 株。在这两个饲养场中，母牛曾接种过含有 A 组 BRV 毒株新生犊牛腹泻病毒（NCDV）-林肯株（P1:G6）的商业疫苗。使用全基因组 RNA 探针进行的 Northern 印迹杂交表明，NS-1 株和 NS-2 株具有相同的 A 组 RNA 电泳图谱，并且在所有基因片段上都是同源的。使用血清学和基因型方法将 NS-1 株与参考 A 组 BRV 毒株进行了比较。体外病毒中和试验表明，NS-1 株可被 G6 特异性中和单克隆抗体（mAb）以及针对 BRV 毒株 B641（P5:G6）产生的豚鼠超免疫血清（GPHS）中和，但不能被 G10 特异性中和 mAb 或针对 BRV 毒株 B223（P11:G10）产生的 GPHS 中和。使用全基因组 RNA 探针进行的核酸杂交实验表明，NS-1 株的基因片段 4 与 BRV 毒株 NCDV-林肯株和 B223 不同，但与 B641 株杂交。相反，NS-1 株的基因片段 5 与 BRV 毒株 B223 杂交，但不与 BRV 毒株 NCDV-林肯株和 B641 杂交。通过逆转录聚合酶链反应（RT-PCR）扩增 NS-1 株产生的 G 特异性 cDNA 探针仅与 G6 毒株 NCDV-林肯株和 B641 特异性杂交，而不与 G10 毒株 B223 杂交。使用 NS-1 株、B641 株和 B223 株进行的共电泳实验进一步证实了这些结果，表明 NS-1 株是 B641 株和 B223 株之间自然发生的重配 BRV。综合这些结果表明，一种自然发生的 A 组 BRV 重配体，其 P 基因与疫苗病毒不同，是导致两个饲养场出现腹泻综合征的原因。本研究结果还表明，感染牛的 A 组 BRV 中至少存在 2 种不同的基因片段 5。
