深圳市首例基孔肯雅病毒的形态学及分子遗传特征分析

目的 对深圳市首发基孔肯雅病毒(CHIKV)进行分离、增殖培养、浓缩、形态学观察及构建系统发生树分析,为后续基因组信息解析、蛋白组分析、单克隆抗体制备及疫苗研发等应用性研究提供实验基础.方法 利用C6/36细胞从病人血清中分离CHIKV,采用BHK-21细胞对CHIKV进行大量的增殖培养,经7％PEG8000浓缩,超薄切片和负染观察CHIKV显微结构;对分离得到的CHIKV株进行全基因组测序和构建系统发生树,结合流行病学资料对其分子遗传特征进行分析.结果 CHIKV被成功分离并浓缩；在透射电镜下观察CHIKV的直径约为70 nm,圆形有包膜,表面有纤突；CHIKV(SZ-20101028)基因组长为12 377bp,属于E1-A226突变株；该病毒是最近10年来在印度洋岛屿爆发流行的新亚型,与流行病学调查资料相吻合.结论 以现有的条件建立了一种稳定、快速培养浓缩CHIKV的方法,并初步对CHIKV进行了形态观察和溯源性分析.

Morphological and Molecular Genetic Characteristics of Chikungunya Virus

Objective To analyze the morphological and genetic characteristics of the chikungunya virus （CHIKV）, and provide basic information for further genomic and proteomic analysis, and vaccine research and development. Methods C6/36 ceils were used for the isolation of CHIKV from patient serum and BHK-21 cells were used for the propagation of the virus. The viral particles were concentrated using 7% PEGSO00. CHIKV microstructure was studied by uhrathin sections and negative staining. Whole genome was sequenced and phylogenetic tree was constructed based on the genome sequence. Results CHIKV was successfully isolated and concentrated. CHIKV （named SZ-20101028） which was isolated from the sera is belong to the El-A226 mutant strain, with a genome size of 12 377bp. The virus is anew subtype of the recent 10 years outbreak in the Indian Ocean islands. Conclusion A new subtype of CHIKV was isolated and propagated in laboratory conditions. The morphology and genome sequence were studied,which provide basic information for further genomic and proteomic analysis, and vaccine research and development.