中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (5): 355-357.

• 研究报道 • 上一篇    下一篇

采用HaCaT细胞培养沙眼衣原体

王蕊1,王敬2,陈立新3,李卓然4,刘原君5,刘全忠6   

  1. 1. 天津医科大学第二医院
    2. 天津医科大学总医院皮肤科
    3. 天津市儿童医院
    4. 天津医科大学第二医院皮肤科
    5. 天津医科大学总医院/天津性传播疾病研究所
    6. 天津医科大学总医院皮肤性病科
  • 收稿日期:2012-08-28 修回日期:2012-11-14 出版日期:2013-05-15 发布日期:2013-05-01
  • 通讯作者: 刘全忠 E-mail:liuquanzhong@medmail.com.cn
  • 基金资助:
    沙眼衣原体临床耐药与体外药敏差异的机制研究及耐药检测方法的优化

Cultivation of Chlamydia trachomatis in HaCaT cells

  • Received:2012-08-28 Revised:2012-11-14 Online:2013-05-15 Published:2013-05-01
  • Contact: quanzhong liu E-mail:liuquanzhong@medmail.com.cn

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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SPSS17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。

关键词: 衣原体，沙眼, 细胞培养, HaCaT 细胞

Abstract: WANG Rui, WANG Jing, CHEN Li-xin, LI Zhuo-ran, LIU Yuan-jun, LIU Quan-zhong *. *Department of Dermatology, General Hospital, Tianjin Medical University, Tianjin 300052, China Corresponding author: LIU Quan-zhong, Email: liuquanzhong@medmail.com.cn 【Abstract】 Objective To investigate the feasibility of C .trachomatis culture in HaCaT human keratinocytes. Methods According to the procedure for C. trachomatis culture in McCoy cells, 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.

Key words: cell culture