中华皮肤科杂志 ›› 1997, Vol. 30 ›› Issue (3): 163-166.

• 论著 • 上一篇    下一篇

用聚合酶链反应和限制性片段长度多态性分析进行沙眼衣原体的基因分型

李珊山, 叶顺章, 王千秋, 钟铭英, 张树文   

  1. 中国医学科学院、中国协和医科大学皮肤病研究所, 南京210042
  • 收稿日期:1995-12-19 修回日期:1996-03-25 出版日期:1997-06-15 发布日期:1997-06-15

Gene Typing of Chlamydia Trachomatis by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism

Li Shanshan, Ye Sunzhang, Wang Qianqiu   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences, Peking Union Medical College, Nanjing 210042
  • Received:1995-12-19 Revised:1996-03-25 Online:1997-06-15 Published:1997-06-15

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摘要： 为了研究泌尿生殖道沙眼衣原体(CT)感染的分子流行病学,我们建立了一种不需做培养的CT血清型鉴定方法。用聚合酶链反应(PCR)扩增CT大部分主要外膜蛋白基因(ompl),将扩增产物做限制性片段长度多态性(RFLP)分析,即用限制性内切酶AluⅠ和MspⅠ同时消化,产生的片段通过10%聚丙烯酰胺凝胶电泳分离。经硝酸银染色后,15个CT血清型标准株产生13个特征性带谱。血清型C和J带型相似,用HinfⅠ酶切后区分;H和L3带型相同,用DdeⅠ和EcoRⅠ同时酶切后区分。对12份CT培养阳性的临床标本行PCR-RFLP分型检测。结果:D血清型为6份标本,E型2份,F型2份,F/G混合型1份,B型1份。研究表明对ompl的PCR-RFLP基因分型方法能用于大范围CT感染的流行病学研究。

关键词: 沙眼衣原体, 外膜蛋白基因, 聚合酶链反应, 限制性片段长度多态性

Abstract: In order to facilitate molecular epidemiologic study of chlamydial urogenital infections,a method that aviods culture was developed to determine the serovars of Chlamydia trachomatis(CT). Polymerase chain reaction was used first to amplify a large part of the major outer membrane protien gene(omp1). The amplified DNA was then digested simultaneously with restriction endonuclease Alu I and Msp I and the resulting fragments were separated on 10% polyacrylamide gels. After silver nitrate staining, a total of 13 characteristic patterns were observed for the 15 serovars. Serovar C and J were similar in pattern, but could be distinguished by cleavage with HinfⅠ. Using Dde I and EcoRI, the patterns of serovar H and L3 could be differentiated from each other. Analysis of 12 CT-positive clinical specimens gave the following results: 6D,2E,2F,1F/G and 1B strains. This study indicates that the omp1-based genotyping is useful for large-scale epidemiologic studies of chlamydial infections.

Key words: Chlamydia trachomatis, Outer memberane protein gene, Polymerase chain reaction, Restriction fragment length polymorphism