The State Key Laboratory of Agricultural Microbiology, Wuhan 430070, China; College of Veterinary Medicine, Cooperative Innovation Centre of Substantial Pig Production, Hubei Province China, Huazhong Agricultural University, Wuhan 430070, China.
Wuhan Keqian Biology Co. Ltd, Wuhan 430023, China.
Infect Genet Evol. 2021 Apr;89:104715. doi: 10.1016/j.meegid.2021.104715. Epub 2021 Jan 9.
Rotaviruses (RVs) account for severe diarrhea in children and young animals globally. In the current study, the fecal samples of diarrheic calves from a beef farm in Inner Mongolia were screened for RVA by ELISA and RT-PCR, followed by culture of three positive RVA samples in the MA-104 cell line. After 10 blind passages, cytopathic effects (CPE) appeared as detachment, granulation, and clustering of the inoculated cells. The virus isolates were identified by RT-PCR (VP6 gene RVA) and ESI-LC-MS/MS for whole protein sequencing. The protein sequences demonstrated the presence of two strains from species A rotavirus and one RVB strain; RVA/Cow-tc/CHN/35333/2019/G6P[5] was mixed with one RVB strain (RVB/Cow-tc/CHN/35334/2019/G5P[3]) in two samples, and RVA/Cow-tc/CHN/10927/2019/G8P[7] was found in one sample. They are of genotype constellations (G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3), (G8-P[7]-I5-R1-C1- M2-A1-N1-T1-E1-H1), and (G5-P[3]-I3-R5-C5-A5-N4-H5), respectively. Besides, phylogenetic analysis of the obtained sequences demonstrated viral evolution.
轮状病毒（RVs）在全球范围内可导致儿童和幼小动物严重腹泻。在本研究中，通过 ELISA 和 RT-PCR 对内蒙古一家肉牛场腹泻犊牛的粪便样本进行了 RVA 筛查，随后将 3 个阳性 RVA 样本在 MA-104 细胞系中进行培养。经过 10 次盲传，接种细胞出现了细胞病变效应（CPE），表现为细胞脱落、颗粒化和聚集。通过 RT-PCR（VP6 基因 RVA）和 ESI-LC-MS/MS 对全蛋白测序对病毒分离物进行了鉴定。蛋白质序列显示存在两种 A 种轮状病毒株和一种 RVB 株；两个样本中 RVA/Cow-tc/CHN/35333/2019/G6P[5]与一种 RVB 株（RVB/Cow-tc/CHN/35334/2019/G5P[3]）混合，一个样本中发现 RVA/Cow-tc/CHN/10927/2019/G8P[7]。它们的基因型组合为（G6-P[5]-I2-R2-C2-M2-A3-N2-T6-E2-H3）、（G8-P[7]-I5-R1-C1-M2-A1-N1-T1-E1-H1）和（G5-P[3]-I3-R5-C5-A5-N4-H5）。此外，获得的序列的系统进化分析表明了病毒的进化。
