采用HaCaT细胞培养沙眼衣原体  

Cultivation of Chlamydia trachomatls in HaCaT cells

作　　者：王蕊[2] 王敬[1] 陈立新[1] 李卓然[3] 刘原君[1] 刘全忠[1] 

机构地区：[1]天津医科大学总医院皮肤科,天津300052 [2]天津医科大学第二医院皮肤科,天津300211 [3]天津医科大学第二医院皮肤科

出　　处：《中华皮肤科杂志》2013年第5期355-357,共3页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30872285）

摘　　要：目的探讨新的培养沙眼衣原体的细胞。方法借鉴McCoy细胞培养沙眼衣原体的方法，将E型标准株和临床标本接种于HaCaT细胞，分别用碘染、单克隆荧光抗体检测、PCR扩增沙眼衣原体内源性质粒的方法检测沙眼衣原体是否可以在体外感染HaCaT细胞。用HaCaT细胞传代培养沙眼衣原体E型标准株5代，分别计数5次传代的包涵体数，并用SPSSl7软件进行单因素方差分析，以确定沙眼衣原体在HaCaT细胞中是否增殖。结果碘染后可见HaCaT细胞胞质内有典型的包涵体。单克隆荧光抗体检测，在荧光显微镜下可见HaCaT细胞内有黄色荧光颗粒。PCR扩增HaCaT细胞内沙眼衣原体内源性质粒为阳性。沙眼衣原体E型标准株在HaCaT细胞内经传代5次，包涵体数逐渐增加。结论HaCaT细胞在体外培养沙眼衣原体成功，且沙眼衣原体E型标准株在HaCaT细胞中可以增殖。Objective To investigate the feasibility of C .trachomat/s culture in HaCaT human keratinocytes. Methods According to the procedure for C. trachomatis culture in McCoy ceils, clinical swab specimens and standard strains of C. trachomatis serotype E were inoculated into HaCaT cells. Iodine staining, a fluorescent monoclonal antibody test and PCR amplification of the endogenous plasmid of C. trachomatis were performed to detect the growth of C. trachomatis in HaCaT cells. Five passages of subculture were carried out for the standard strain of C. trachomatis serotype E in HaCaT cells, and inclusion bodies were counted after each passage. One-factor analysis of variance was conducted by using the software SPSS17 to determine if C. trachomatis was propagated in HaCaT cells. Results Iodine staining showed typical inclusion bodies of C. trachomatis in the cytoplasm of HaCaT cells. Yellow fluorescence-labeled granules were observed in the HaCaT cells under a microscope. Endogenous plasmids of C. rachomatis were successfully amplified by PCR from the infected HaCaT cells. The number of inclusions in HaCaT cells gradually increased at passage 1 through 5. Conclusions C. trachomatis is successfully cultivated in HaCaT cells in vitro, and the standard strain of C. trachomatis serotype E can propagate in HaCaT cells.

关 键 词：沙眼衣原体 T细胞培养 HACAT细胞 荧光抗体检测 单因素方差分析 PCR扩增 细胞传代培养 MCCOY 

分 类 号：R3[医药卫生—基础医学]