Parwani A V, Hussein H A, Rosen B I, Lucchelli A, Navarro L, Saif L J
Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Wooster.
J Clin Microbiol. 1993 Aug;31(8):2010-5. doi: 10.1128/jcm.31.8.2010-2015.1993.
Dot and Northern blot hybridization assays were used to analyze field strains of group A bovine rotaviruses (BRVs) by using nucleic acid probes representing P and G type specificities. The probes were prepared by polymerase chain reaction amplification of hyperdivergent regions of the cloned VP4 (nucleotides 211 to 686) and VP7 (nucleotides 51 to 392) genes from four serotypically distinct (in P or G types) strains of rotaviruses: NCDV (G6, P1), IND (G6, P5), 69M (G8, P10), and Cr (G10, P11). The P and G type cDNA probes were radiolabeled with [32P]dCTP and hybridized with RNA extracted from reference cell culture-passaged rotavirus strains or the field samples. The field samples were obtained from young diarrheic calves from Ohio, Nebraska, Washington State, and Canada. The cDNA probes were specific for their respective G or P types on the basis of analysis of known P and G type reference strains. The G typing analysis of 102 field samples revealed that 36.3% (37 of 102) were G6, 2.9% (3 of 102) were G8, 12.7% (13 of 102) were G10, and 23.5% (24 of 102) were untypeable. The P typing results for 93 samples indicated that 2.2% (2 of 93) were P1 (NCDV-like), 20.4% (19 of 93) were P5 (UK-like), 9.3% (10 of 93) were P11 (B223-like), and 40.8% (38 of 93) were untypeable. This is the first report of the identification among BRV strains in North America of a G type other than G6 or G10. Our report further confirms that G6, P5 rotaviruses are predominant among the BRV field strains that we examined, and the P types of these strains differ from that of the BRV vaccine strain used in the United States (G6, P1). The large number of untypeable G (23.5%) and P (40.8%) types suggests that other or new P and G types exist among BRV field strains.
采用斑点杂交和Northern杂交分析方法，利用代表P型和G型特异性的核酸探针，对A组牛轮状病毒（BRV）的野毒株进行分析。这些探针通过聚合酶链反应扩增轮状病毒4个血清型不同（P型或G型）毒株（NCDV，G6、P1；IND，G6、P5；69M，G8、P10；Cr，G10、P11）的克隆VP4基因（核苷酸211至686）和VP7基因（核苷酸51至392）的高变区制备而成。P型和G型cDNA探针用[32P]dCTP进行放射性标记，并与从参考细胞培养传代的轮状病毒毒株或野毒株样本中提取的RNA杂交。野毒株样本取自俄亥俄州、内布拉斯加州、华盛顿州的腹泻幼牛以及加拿大的腹泻幼牛。根据对已知P型和G型参考毒株的分析，cDNA探针对其各自的G型或P型具有特异性。对102份野毒株样本的G型分析显示，36.3%（102份中的37份）为G6型，2.9%（102份中的3份）为G8型，12.7%（102份中的13份）为G10型，23.5%（102份中的24份）无法分型。对93份样本的P型分析结果表明，2.2%（93份中的2份）为P1型（类似NCDV），20.4%（93份中的19份）为P5型（类似UK），9.3%（93份中的10份）为P11型（类似B223），40.8%（93份中的38份）无法分型。这是北美地区在BRV毒株中鉴定出G6或G10以外G型的首次报道。我们的报告进一步证实，G6、P5轮状病毒在我们检测的BRV野毒株中占主导地位，并且这些毒株的P型与美国使用的BRV疫苗毒株（G6、P1）不同。大量无法分型的G型（23.5%）和P型（40.8%）表明，BRV野毒株中存在其他或新的P型和G型。
