中华皮肤科杂志

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沙眼衣原体omp1基因VS1-VS2序列分析与基因分型

郑和平1, 江丽芳1, 方丹云1, 吴兴中2, 李燕娃3, 许刚4, 薛耀华2, 冯占芹2, 吴亚安2, 黄进梅2   

  1. 1. 中山大学中山医学院, 广州510089;
    2. 广东省皮肤性病防治中心;
    3. 佛山市皮肤病防治所;
    4. 珠海市慢性病防治所
  • 收稿日期:2006-04-18 出版日期:2007-05-15 发布日期:2007-05-15
  • 通讯作者: 江丽芳,email:jianglf909@yahoo.com.cn E-mail:jianglf909@yahoo.com.cn
  • 基金资助:
    广东省科委资助项目(2005B34201011);广东省卫生厅资助项目(A2005150)

Genotyping of Chlamydia trachomatis by sequence analysis of VS1-VS2 region of omp1 gene

ZHENG He-ping1, JIANG Li-fang1, FANG Dan-yun1, WU Xing-zhong2, LI Yan-wa3, XU Gang4, XUE Yao-hua2, FENG Zhan-qin2, WU Ya-an2, HUANG Jin-mei2   

  1. Department of Microbiology, School of Medicine, Sun Yat-Sen University, Guangzhou 510089, China
  • Received:2006-04-18 Online:2007-05-15 Published:2007-05-15

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摘要： 目的 探讨沙眼衣原体omp1基因VS1-VS2序列基因分型和VS1-VS2基因突变率。方法 巢式PCR扩增omp1基因的VS1-VS2序列,自动测序仪测序,运用DNAstar软件进行临床菌株与标准菌株序列的比对。结果 99株临床标本分离株扩增出约453 bp大小的VS1-VS2片段。与标准株比对,除3株测序出现双峰无法确定基因型外,96株菌株分为9个型,以F型29株(30.2%)、E型22株(22.9%)、J型19株(19.8%)、D型12侏(12.5%)和H型8株(8.3%)为主。序列分析发现7株存在VS1区碱基的突变或插入,多为有义突变,突变率为7.3%,多见于D、E、F和H型。结论 沙眼衣原体omp1基因序列分析在分子流行病学上具有一定意义。

关键词: 衣原体, 沙眼, 序列分析, DNA, 基因型, 突变

Abstract: Objective To type clinical strains of C.trachomatis by sequence analysis of VS1-VS2 region of omp1 gene,and to investigate the mutation rate of this gene.Methods Urogenital specimens were collected from 99 patients attending our STD clinic.The VS1-VS2 region of omp1 gene was amplified in these specimens by nested PCR,and sequenced with an automatic sequencer.The sequencing results were blasted against sequences of reference strains by DNAstar software.Results A 435-bp fragment of VS1-VS2 region was amplified from all 99 clinical samples.Three specimens appeared to contain multiple pathogens and could not be genotyped.The remaining 96 strains were classified into 9 genotypes.The most prevalent types in decreasing order were F(29,30.2%),E(22,22.9%),J(19,19.8%),D(12, 12.5%)and H(8,8.3%).Seven(7.3%)strains showed base mutation or insertion in VS1 region,most of which were sense mutation and primarily occurred among genotypes D,E,F and H.Conclusion Sequencing analysis of omp1 gene may be useful in the molecular epidemiology of C.trachomatis infection.

Key words: Chlamydia trachomatis, Sequence analysis,DNA, Genotype, Mutation