中华皮肤科杂志 ›› 2016, Vol. 49 ›› Issue (11): 796-800.

• 论著 • 上一篇    下一篇

湖北地区须毛癣菌复合体的分子鉴定

陈瑶,董碧麟,童中胜,李东升   

  1. 武汉市第一医院皮肤科
  • 收稿日期:2016-03-23 修回日期:2016-06-15 出版日期:2016-11-15 发布日期:2016-10-28
  • 通讯作者: 李东升 E-mail:LDS1204@126.com

Molecular identification of Trichophyton mentagrophytes complex strains in Hubei province

  • Received:2016-03-23 Revised:2016-06-15 Online:2016-11-15 Published:2016-10-28
  • Contact: LI DongSheng E-mail:LDS1204@126.com

摘要:

目的 对分离自患者不同部位的须毛癣菌临床株进行种内分型研究,比较不同靶位对须毛癣菌复合体的鉴定效能。方法 选取武汉市第一医院皮肤科近3年临床分离的须毛癣菌48株,经形态学观察和尿素酶试验进行表型鉴定。分别以核糖体DNA(rDNA)的转录间隔区(ITS)和核糖体大亚基(28S rDNA)D1?D2区为靶位,PCR扩增ITS区和D1?D2区后对产物进行测序,联合应用Clustal X2和MEGA 6.0软件,采用最大似然度法进行序列分析并建立遗传进化树。结果 ITS树提示,受试菌株和趾间毛癣菌聚类在同一主支的概率为92%,D1?D2树无法有效区分癣毛癣菌昆克变种和趾间毛癣菌。趾间毛癣菌形态多变,可表现为羊毛型、绒毛型、乳皮型、粉末型和颗粒型。结论 ITS区可将须毛癣菌鉴定到种水平,其鉴定效能优于D1?D2区,形态学方法不能作为其种内鉴定的依据。

Abstract:

Chen Yao, Dong Bilin, Tong Zhongsheng, Li Dongsheng Department of Dermatology, Wuhan No. 1 Hospital, Wuhan 430022, China Corresponding author: Li Dongsheng, Email: LDS1204@126.com 【Abstract】 Objective To profile the intraspecific type of Trichophyton mentagrophytes clinically isolated from different anatomical sites of patients, and to compare the performance of different target sites for the identification of Trichophyton mentagrophytes complex strains. Methods A total of 48 Trichophyton mentagrophytes strains, which were clinically isolated from Department of Dermatology, Wuhan No. 1 Hospital in the latest 3 years, were included in this study. The phenotypes of these Trichophyton mentagrophytes isolates were primarily determined by morphological observation and the urease test. PCR was performed to amplify the nuclear ribosomal internal transcribed spacer (ITS) region and the D1-D2 domains of the large-subunit ribosomal DNA (28S rDNA) followed by DNA sequencing. Then, Clustal X2 software and MEGA 6.0 software were used to analyze the ITS and D1-D2 sequences and to build phylogenetic trees by the maximum-likelihood method (bootstrap = 2000). Results As the ITS sequence-based phylogenetic tree showed, the probability that the 48 isolates were grouped into the Trichophyton interdigitale clade reached 92%. However, Trichophyton interdigitale could not be effectively differentiated from Trichophyton quinckeanum by the D1-D2 sequence-based phylogenetic tree. In addition, Trichophyton interdigitale showed various appearances, including woolen type, downy type, cream type, powdery type and granular type. Conclusions Trichophyton mentagrophytes can be identified to the species level based on the sequence of ITS region, which shows higher efficiency in identifying Trichophyton mentagrophytes complex than the D1-D2 domains. Morphological characteristics can not serve as the basis for intraspecific typing of Trichophyton mentagrophytes.