关键词: 树鼩, 轮状病毒, 反转录聚合酶链式反应, 细胞培养

Abstract: Objective　Isolate and identify the virus from the feces of wild tree shrews, and explore the establishment of method for isolating and cultivating tree shrew rotaviruses.Methods　Fecal samples from tree shrews collected in the wild were used to amplify pathogen RNA via reverse transcription polymerase chain reaction (RT-PCR) using specific primers. The amplified products were separated by agarose gel electrophoresis, and the target bands were purified through gel extraction for subsequent analysis.Results The target fragment of 365 bp was obtained via RT-PCR. After sequencing, the gene fragment showed a nucleic acid sequence coverage of 92% and a matching consistency of 99% when compared with the VP7 segment sequence of bovine rotavirus on the Basic Local Alignment Search Tool (BLAST) in the National Center for Biotechnology Information (NCBI) database. Significant cytopathic effects (CPE) were observed upon inoculation into normal MA104 cells.Conclusion　The rotavirus strain carried by the tree shrew isolated in this study has extremely high sequence homology with the rotavirus strain carried by mammalian cattle (coverage 92%, base identity 99%), which has certain reference value for the isolation and identification of tree shrew rotavirus in the future.

Key words: tree shrew, rotavirus, reverse transcription polymerase chain reaction, cell culture