基于omp1基因VS1-VS2序列的沙眼衣原体RFLP基因分型  

Combination of PCR Targeting the VS1-VS2 of omp1 and Restriction Fragment Length Polymorphism(RFLP) Analysis for Genotyping of Urogenital Chlamydia Trachomatis

作　　者：吴亚安[1] 郑和平[2] 吴志华[1] 李燕娃 吴兴中[2] 

机构地区：[1]广东医学院,广东湛江524023 [2]广东省皮肤性病防治中心,广东广州510500 [3]广东省佛山市皮肤病防治所,广东佛山528000

出　　处：《岭南皮肤性病科杂志》2007年第2期73-75,共3页Southern China Journal of Dermato-Venereology

摘　　要：目的:建立和评价基于omp1基因VS1-VS2序列的沙眼衣原体RFLP基因分型技术。方法:采用巢式聚合酶链反应(PCR)扩增omp1基因的VS1-VS2序列,以限制性酶切片段多态性分析(RFLP)进行检测分型,与VS1-VS2测序分型结果比较。结果:扩增12株沙眼衣原体标准株omp1基因VS1-VS2序列,长度为453-463 bp,用AluⅠ及DdeⅠ酶切此得出标准血清型的特征性图谱。37例沙眼衣原体ELISA检测阳性标本的VS1-VS2-PCR扩增均阳性,RFLP检到B-K型和2例混合型感染,分型率为94.59%。与DNA直接基因测序分型进行对比,符合率为94.59%。结论:基于omp1基因VS1-VS2序列的沙眼衣原体RFLP基因分型技术,较以前的方法更为敏感和特异,可作为对沙眼衣原体分型研究的工具。Objective：To develop and evaluate a new PCR-based assay targeting the VS1-VS2 region of the ompl gene for detecting and typing of urogenital chlamydia trachomatis. Methods： The VS1-VS2 of the ompl gene was amplified by the nested PCR and restriction fragment length polymorphism （RFLP） assay was used to genotype chlamydia trachomatis. The results by RFLP were compared with the genotyps by sequencing the VS1-VS2 region of the omp1 gene of chlamydia trachomatis. Results： 453-463 bp fragments of VS1-VS2 were amplified from 12 referent strains with different serovars. Characteristic patterns of RFLP for the referent strains were obtained by digesting the PCR products by Alu I or Dde I . Thirty seven ELISA-positive samples were amplified and 94.59% （35/37）strains could be genotyped. A total of eight genotypes （B to K） were identified and 94. 59% concordance for all of 37 clinical samples were founded by comparison with the genotypes by sequencing. Conclusion： This study showed that the RFLP assay targeting the VS1-VS2 of ompl are feasible for directly detecting and genotyping of chlamydia trachomatis in urogential samples. It is sensitive and specific, and useful for epidemiological study of chlamydia trachomatis infection.

关 键 词：沙眼衣原体 巢式聚合酶链反应 基因分型 

分 类 号：R374[医药卫生—病原生物学]