Cloning of Omp1 Gene from Chlamydia trachomatis F Genotype

Objective: To directionally clone the ompl gene fromChlamydia trachomatis (Ct) F genotype onto a plasmid vectorfor constructing a rudimentary DNA vaccine. Methods: The complete ompl gene from genomic DNA of CtF genotype wild species was amplified with primers designedby computer. The recombinant gene was obtained byrestriction enzyme cutting, linking the gene with the plasmidvector in vitro, transforming the recombinant gene intobacteria, and extracting the DNA from the bacteria. Results: DNA extracted from the bacteria was composed ofthe ompl gene and plasmid, which is identified by threemethods of singular restrictive enzyme cutting, doublerestrictive enzyme cutting and PCR. Conclusion: Cloning of the ompl gene from the Ct Fgenotype means that a rudimentary DNA vaccine wassuccessfully constructed.

基于Omp1基因的泌尿生殖道感染沙眼衣原体多样性及分布特征分析

目的基于主要外膜蛋白基因(Omp1)研究泌尿生殖道感染患者沙眼衣原体多态性及分布特征,为该疾病的诊治及疫苗研究奠定基础。方法对泌尿生殖道感染临床标本常规处理后,使用沙眼衣原体核酸检测试剂盒筛选阳性标本,然后以巢式PCR扩增Omp1基因,选择4个限制性内切酶进行RFLP分析确定其基因型,选择代表菌株测定Omp1的DNA序列并与参比菌株比较确定其血清型,最后将基因型及血清型与患者的性别、科室及年龄进行对比分析,研究其分布特征。结果 2013-2014年2年间共筛选得到178份阳性标本,61份标本扩增获得Omp1基因,Omp1-PFLP分析将61份标本分为6个基因型,占比分别为24.59%,24.59%,4.92%,9.84%,3.28%和32.79%,血清型分析表明61份标本属于F,H,J,G,Da,D和E 7种血清型,E型及F型占比最多。结论本地区引起泌尿生殖道感染的沙眼衣原体主要属于F,H,J,G,Da,D和E 7种血清型,E型及F型占比最多,基因型及血清型与患者性别具有相关性,与科室、年龄无相关性。

湘西地区女性泌尿生殖道沙眼衣原体感染状况及基因分型初步研究

目的对湘西地区女性泌尿生殖道沙眼衣原体的感染状况及基因分型进行初步研究。方法利用单克隆抗体直接免疫荧光法(DFA)对100例宫颈病变患者及77位健康女性进行筛查,对沙眼衣原体感染者的宫颈管内拭子标本直接进行套式PCR,扩增主要外膜蛋白基因(Omp1)第4可变区(VS4),对将扩增的Omp1VS4基因片段进行测序、分型,并进行同源性分析。结果DFA检测177名妇女的宫颈拭子标本,36份沙眼衣原体阳性,29份阳性标本扩增出277bp目的DNA。经基因测序,共检出8个基因型,其中E型9株(31.0%)、F型7株(24.1%)、G型5株(17.2%)、D型3株(10.3%)、H型2株(6.9%)、J型1株(3.5%)、Ba型1株(3.5%)、K型1株(3.5%),4例存在Omp1基因变异。结论湘西地区女性存在生殖道沙眼衣原体感染,利用对基因的多态性分析可以对沙眼衣原体感染流行情况作出分析。