中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (9): 646-649.

• 论著 • 上一篇    下一篇

荧光原位杂交法观察小鼠皮肤组织切片中型无绿藻感染的研究

康俞莉1,赵颖2,章强强3   

  1. 1. 复旦大学附属华山医院
    2. 上海 复旦大学附属华山医院皮肤科
    3. 上海复旦大学华山医院皮肤科
  • 收稿日期:2013-12-11 修回日期:2014-01-15 出版日期:2014-09-15 发布日期:2014-09-01
  • 通讯作者: 章强强 E-mail:zhangqq8@163.com

Detection of Prototheca zopfii infection in mouse skin tissue sections by using fluorescence in situ hybridization

  • Received:2013-12-11 Revised:2014-01-15 Online:2014-09-15 Published:2014-09-01
  • Contact: Qiangqiang Zhang E-mail:zhangqq8@163.com

摘要: 目的 探讨荧光原位杂交技术用于诊断小鼠皮肤中型无绿藻感染模型的可行性。 方法 制作小鼠皮肤中型无绿藻感染模型,制备皮肤组织石蜡切片,用人工合成的探针PZ-probe进行荧光原位杂交检测,以过碘酸锡夫(PAS)染色、HE染色做阳性对照;取正常及其他真菌感染的皮肤组织石蜡切片做阴性对照,以同样的方法进行荧光原位杂交检测,将各种检测方法的结果进行对比分析。 结果 小鼠临床体征、病理检测及病原体培养结果均证实皮肤无绿藻感染模型建立成功。通过荧光原位杂交法成功检测出了小鼠皮肤中型无绿藻感染模型皮肤组织中的无绿藻病原体,结果与PAS染色和HE染色一致;而在正常及其他真菌感染的皮肤组织石蜡切片中均未能检测出无绿藻。 结论 荧光原位杂交法可以检测出小鼠皮肤中型无绿藻感染模型组织石蜡切片中的无绿藻病原体。

关键词: 原壁菌属, 皮肤, 原位杂交,荧光, 小鼠

Abstract: Kang Yuli, Zhao Ying, Zhang Qiangqiang. Department of Dermatology, Huashan Hospital, Fudan University, Shanghai 200040, China Corresponding author: Zhang Qiangqiang, Email: zhangqq8@163.com 【Abstract】 Objective To evaluate the feasibility to detect Prototheca in a mouse model of Prototheca zopfii cutaneous infection by using fluorescence in situ hybridization (FISH). Methods The model of Prototheca zopfii cutaneous infection was established by abdominal subcutaneous inoculation of Prototheca zopfii suspensions into 20 male BALB/c mice. Seven days after the inoculation, the mice were sacrificed, and tissue specimens were obtained from abdominal skin and subjected to microscopic examination, fungal culture and paraffin embedding. A PZ-probe was artificially synthesized and used to detect Prototheca in paraffin-embedded sections by using FISH. Moreover, both periodic acid-Schiff (PAS) and hematoxylin-eosin (HE) staining were performed to examine the paraffin-embedded sections. Skin specimens obtained from normal mice and Candida albicans- or Cryptococcus neoformans-infected mice served as the negative control. Results Clinical presentations, pathological examination and fungal culture results all confirmed the successful establishment of Prototheca zopfii skin infection model in mice. Prototheca was identified by FISH with the PZ-probe in the paraffin-embedded skin tissue sections from the murine model of Prototheca zopfii cutaneous infection, but not detected in the negative control tissue specimens, which was consistent with the results of PAS and HE staining. Conclusion FISH can be used to detect Prototheca in paraffin-embedded skin sections from the mouse model of Prototheca zopfii cutaneous infection.

Key words: Prototheca, Skin, In situ hybridization, fluorescence, Mice

中图分类号: 

  • R756.9