猴腺病毒GD株的分离与鉴定

目的 体外分离出猴腺病毒(simian adenovirus,SAdV)毒株,并对其进行鉴定,为SAdV的生物学特性、致病机制和诊断试剂等方面的研究提供基础.方法 采用PCR法对猕猴粪便样本进行SAdV筛查,将PCR检测为阳性的样本处理后接种BSC-1细胞,盲传3代后用PCR扩增测序并负染色电镜观察.结果 从广东地区食蟹猴粪便样本中成功分离出一株SAdV,细胞病变明显,PCR检测呈阳性,电镜下可观察到典型腺病毒样形态的病毒粒子,命名为GD株.经测序鉴定,发现GD株属HAdV-G亚属,与SAdV-1进化发生距离最近.结论 GD株的分离成功,使猴腺病毒基因转移载体的构建成为可能,有助于猴腺病毒替代载体的发展.

Isolation and identification of simian adenovirus strain GD

Objective To isolate simian adenovirus（SAdV） in vitro.Methods Simian adenovirus in fecal specimens of monkeys were detected by PCR.SAdV-positive samples were inoculated into BSC-1 cells.The cells were blindly passaged for three times to observe cytopathic effect in cell cultures.The presence of SAdV was confirmed by PCR-sequencing and electron microscopy.Results A strain of monkey adenovirus was isolated form Macaca fascicularis in Guangdong province,hence it was named as GD strain.The cytopathic effect was obvious 24 hours after infection.Viral particle-like adenovirus were observed by electron microscopy.The GD strain had high nucleotide sequence homology with HAdV-G.Pol and hexon genes of the isolated strain had 96% and 83% nucleotide sequence homology with SAdV-1,respectively.Conclusions The successful isolation of SAdV can facilitate the research for its biology,pathogenesis and laboratory diagnostics,and make it possible to construct SAdV gene transfer vector that benefits development of alternative carrier.