关键词: 犬蝠, 呼肠病毒, 分离, 鉴定

Abstract: Wild short-nosed fruit bats samples were collected from Shaoguan City Guangdong Province. The samples were scrunched and infected Vero-E6 cells. Two strains viruses were isolated. One virus isolated from short-nosed fruit bats hadn’t caused cytopathic effects (CPE) until fourth-passage on Vero-E6 cells. Infected cells emerged granulating, shrinking, rounding and falling off. After thrice freeze-thaw cells and culture medium were harvested for electron microscopy. Virus particles were nonenveloped, double capsids and icosahedral symmetry. The virus was denominated Bat/China/2003(B/03). Hemagglutination test indicated this virus could agglutinate healthy human type O red cells, but could not agglutinate SPF chicken, experimental common bovine, rat and guinea pig red cells. Nucleic acid electrophoresed on agarose gels suggested that the genome of this virus was divided into large, medium, small three parts. One pairs of specific primers according to mammalian reovirus were used for Reverse Transcription Polymerase Chain Reaction (RT-PCR). Appropriate specific products were amplified by RT-PCR. NCBI BLAST analysis indicated that this segment shares the highest identity with Ndelle virus which is a member of the genus Orthoreovirus, and the homology is 91.2 %. DNAMAN Multiple Alignment analysis of nucleotide sequences with mammalian reovirus serotype 1(T1L), 2(T2J) and 3(T3D). Homologies were 89.9%, 76.9% and 89.9%, respectively. So we can deduce this virus is a member of the virus family Reoviridae.

Key words: Short-nosed fruit bat, Reovirus, Isolation, Identification