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中国真菌学杂志 2011, Vol. 6  Issue (2): 70-76.

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我国代表地区须癣毛癣菌复合体的分子鉴定与分型研究

胡小平, 万喆, 王晓红, 李若瑜   

  1. 北京大学第一医院皮肤性病科真菌室, 北京大学真菌和真菌病研究中心, 北京, 100034
  • 收稿日期:2010-12-03 出版日期:2011-04-28 发布日期:2011-04-28
  • 通讯作者: 李若瑜,E-mail:lry0660@gmail.com E-mail:lry0660@gmail.com
  • 作者简介:胡小平,男(汉族),硕士,主治医师.E-mail:xiaoping7752@sohu.com
  • 基金资助:
    卫生部公益项目重点基金(200802026);科技部十一五传染病重大专项课题(20082x10004-002)

Study on molecular identification and typing of Trichophytonmentagrophytes complex from the representative regions in China

HU Xiao-ping, WAN Zhe, WANG Xiao-hong, LI Ruo-yu   

  1. Departmemt of Dermatovenerology, Peking University First Hospital, and Research Center for Medical Mycology, Beijing 100034 China
  • Received:2010-12-03 Online:2011-04-28 Published:2011-04-28

摘要: 目的 对我国代表地区的须癣毛癣菌菌株进行分子再鉴定和分型研究。方法 选取我国南北方8个省市地区经表型鉴定的须癣毛癣菌菌株47株,通过再培养形态观察、生理试验;PCR扩增核糖体DNA(rDNA)的内转录间隔区(ITS)和核糖体大亚基(LSU)D1-D2区,测序后利用数据库进行序列比对,对须癣毛癣菌复合体进行再鉴定;PCR扩增rDNA非转录间隔区(NTS)的三个串联重复亚单位S0、S1和S2区,进行种内分型,并比较不同部位来源菌株型别的差异性。结果 我国南北方8个省市地区47株须癣毛癣菌中3株鉴定为断发毛癣菌,6株鉴定为无性型苯海姆节皮菌,其余均鉴定为万博节皮菌中的亲人型趾间毛癣菌;三对不同引物扩增38株趾间型毛癣菌和2株苯海姆节皮菌NTS区,共产生28种特征性带型。带型和菌株来源及发生部位无相关性。结论 我国分离自人类须癣毛癣菌复合体的主要组成菌种为趾间毛癣菌;ITS区结合LSU D1-D2区测序有助于鉴定须癣毛癣菌复合体至种水平;NTS区的三个串联重复亚单位所产生的特征性指纹图提供了一种快速、稳定的分子生物学种内分型方法,可应用于趾间毛癣菌感染的流行病学研究。

关键词: 须癣毛癣菌复合体, rDNA内转录间隔区, 核糖体大亚基D1-D2区, rDNA非转录间隔区

Abstract: Objective To study molecular identification and typing of Trichophyton mentagrophytes complex from the representative regions in China.Methods A total of 47 strains of Trichophyton mentagrophytes(T.mentagrophytes) identified by phynotype characteristic from 8 different regions of China were re-identified by sequence analysis of the rRNA gene internal transcribed spacer regions(ITS) and the D1-D2 domain of the large-subunit rRNA gene(LSU D1-D2),and typed by PCR fingerprinting analysis of Trichophyton mentagrophytes using polymorphic subrepeat loci in the rDNA nontranscribed spacer(NTS).The relationship between type and origin of strains was studied.Results Three strains of Trichophyton tonsurans,6 strains of A.benhamiae,and 38 strains of T.interdigitale were identified.Combining PCR fingerprints from each of the three polymorphic loci produced a total of 28 individual strain profiles.No relationship was observed between profiles and the origin and location of strains.Conclusions Trichophyton interdigitale was the most common species from human beings in China.Sequence analysis of ITS combining LSU D1-D2 arehelpful for identification and the typing method using NTS is rapid,reproducible and discriminatory for fragments interpreting.PCR fingerprint analysis of variable tandem repeat loci in the T.mentagrophytes NTS is valuable for future epidemiological investigations.

Key words: Trichophyton mentagrophytes complex, internal transcribed spacer, D1-D2 domain of the large-subunit rRNA gene, nontranscribed spacer

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