Chang K O, Parwani A V, Saif L J
Department of Veterinary Preventitive Medicine, Ohio Agricultural Research and Development Center, Ohio State University, Wooster, USA.
Arch Virol. 1996;141(9):1727-39. doi: 10.1007/BF01718295.
Characterization of the VP7 (G type) and VP4 (P type) genes of bovine group A rotaviruses (BRV) from field samples was performed using RT-PCR and RFLP analysis. After RT-PCR amplification of the full length VP7 genes and partial length VP4 genes (nucleotides 1 to 1096), four enzymes, EcoRV, NlaIV, BamHI and HpaII were used for digestion analysis. For VP7, four RFLP profiles were observed after analysis of the digests: they were designated as G6, G6s (subtype, showed about 86% nucleotide and 90% amino acid identity to reference G6 strains), G8 and G10. For VP4, three RFLP profiles were observed: designated as P[1], P[5] and P[11]. The G typing analysis of 86 BRV fecal samples from 5 states, representing at least 11 different herds revealed that 60.5% (52/86) were G6, which included G6s (9/52); 19.8% (17/86) were G10; 7% (6/86) were G8; 10.4% (9/86) were G6 and G10 mixtures including two G6s samples; and 2.3% (2/86) were G6 and G6s mixtures. The P typing analysis of the same 86 fecal samples revealed that 64% (55/86) were P[5]; 28% (24/86) were P[11]; 1.2% (1/86) were P[1] and 6 samples (7%) were mixtures of either P[11] or P[5]. When the same samples were analyzed according to G and P type specificity, all possible combinations of G and P types existed in the field. The G6P[5] type was most prevalent and accounted for 46.7% (41/86) of the samples; 12.8% (11/86) were G10P[11]; 7% (6/86) were G10P[5] and an equal number were G6sP[11]. The G6P[11] (n = 2), G8P[1] (n = 1), G8P[5] (n = 1) and G8P[11] (n = 3) combinations were also observed. The following mixed BRV infections were observed in the field samples; G6sP[5 + 11] (n = 1), G8P[5 + 11] (n = 1), G6 + G10P[5] (n = 1) G6 + G10P[5 + 11] (n = 2), G6 + G6sP[11] (n = 1), G6 + G6sP[1 + 11] (n = 1), G6s + G10P[11] (n = 1) and G6s + G10P[5 + 11] (n = 1). Information on the G and P types and G/P combinations in the field samples should be useful for understanding the epidemiology of BRV and designing vaccination strategies to control BRV in the field.
利用逆转录聚合酶链反应（RT-PCR）和限制性片段长度多态性分析（RFLP）对来自现场样本的A组牛轮状病毒（BRV）的VP7（G型）和VP4（P型）基因进行了特征分析。在对全长VP7基因和部分长度VP4基因（核苷酸1至1096）进行RT-PCR扩增后，使用四种酶，即EcoRV、NlaIV、BamHI和HpaII进行消化分析。对于VP7，分析消化产物后观察到四种RFLP图谱：它们被指定为G6、G6s（亚型，与参考G6毒株显示约86%的核苷酸和90%的氨基酸同一性）、G8和G10。对于VP4，观察到三种RFLP图谱：指定为P[1]、P[5]和P[11]。对来自5个州的86份BRV粪便样本进行G分型分析，这些样本代表至少11个不同的牛群，结果显示60.5%（52/86）为G6，其中包括G6s（9/52）；19.8%（17/86）为G10；7%（6/86）为G8；10.4%（9/86）为G6和G10混合物，包括两份G6s样本；2.3%（2/86）为G6和G6s混合物。对相同的86份粪便样本进行P分型分析，结果显示64%（55/86）为P[5]；28%（24/86）为P[11]；1.2%（1/86）为P[1]，6份样本（7%）为P[11]或P[5]的混合物。当根据G和P型特异性对相同样本进行分析时，现场存在所有可能的G和P型组合。G6P[5]型最为普遍，占样本的46.7%（41/86）；12.8%（11/86）为G10P[11]；7%（6/86）为G10P[5]，数量相同的为G6sP[11]。还观察到G6P[11]（n = 2）、G8P[1]（n = 1）、G8P[5]（n = 1）和G8P[11]（n = 3）组合。在现场样本中观察到以下混合BRV感染；G6sP[5 + 11]（n = 1）、G8P[5 + 11]（n = 1）、G6 + G10P[5]（n = 1）、G6 + G10P[5 + 11]（n = 2）、G6 + G6sP[11]（n = 1）、G6 + G6sP[1 + 11]（n = 1）、G6s + G10P[11]（n = 1）和G6s + G10P[5 + 11]（n = 1）。现场样本中关于G和P型以及G/P组合的信息对于了解BRV的流行病学和设计控制现场BRV的疫苗接种策略应该是有用的。
