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目的探讨泌尿生殖道感染沙眼衣原体PmpF基因的多态性及与临床感染的关系。方法收集2013年1月到2014年12月于解放军252医院就诊患者送检的泌尿生殖道分泌物标本,标本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PmpF基因,扩增得到的PmpF基因选用4种内切酶进行酶切及RFLP分析并确定其基因型,根据PmpF-RFLP结果,选择代表菌株测定PmpF基因序列进行系统发育分析,进一步验证PmpF-RFLP结果,同时测定代表菌株的Omp1基因序列并与参考菌株进行比较,从而确定代表菌株的血清型。通过χ~2检验探讨沙眼衣原体PmpF基因型与患者性别、年龄的关系以反映其临床分布特征。结果 2年间从妇科、泌尿科和皮肤科3个科室获得阳性标本178份,117份标本扩增得到PmpF基因,PmpF-RFLP将117份标本分为3个Type型,占比分别为2.6%、36.8%、60.7%,挑选26株菌株进行Omp1序列测定,结果显示26株菌株属于D、Da、E、F、G、H、J 7种血清型,统计学分析表明PmpF基因型与患者性别、年龄的Fisher精确检验值分别为0.167、0.008。结论本地区引起泌尿生殖道感染的沙眼衣原体可以根据PmpF基因分成3个基因型,包括D、Da、E、F、G、H、J 7种血清型,血清型与PmpF基因型无对应关系,PmpF基因型与患者年龄有关,与性别无关。
Objective To investigate the polymorphism of PmpF gene in genitourinary tract infected with Chlamydia trachomatis and its relationship with clinical infection. Methods Genitourinary secretions were collected from patients who visited the People’s Liberation Army at the 252 Hospital from January 2013 to December 2014. Positive samples were screened by using the C. trachomatis nucleic acid detection kit after the samples were treated. Then the PmpF gene was amplified by nested PCR According to the result of PmpF-RFLP, we chose the representative strains to determine the sequence of PmpF gene for phylogenetic analysis, and further verified the results of PmpF-RFLP by PmpF-RFLP. The Omp1 gene sequences of the representative strains were also determined and compared with the reference strains to determine the serotypes of the representative strains. The relationship between PmpF genotype of Chlamydia trachomatis and gender and age of patients was analyzed by χ ~ 2 test to reflect the clinical distribution characteristics. Results 178 positive samples were obtained from 3 departments of gynecology, urology and dermatology in 2 years. PmpF gene was amplified from 117 samples, and 117 samples were divided into 3 types by PmpF-RFLP, accounting for 2.6% 36.8% and 60.7% respectively. Twenty-six isolates were selected for Omp1 sequencing. The results showed that 26 isolates belonged to D, Da, E, F, G, H and J serogroups. Statistical analysis showed that PmpF genotype was positively correlated with sex, Fisher’s exact test values for age were 0.167 and 0.008 respectively. Conclusion Chlamydia trachomatis causing urogenital tract infection in this region can be divided into 3 genotypes according to PmpF gene, including 7 serotypes of D, Da, E, F, G, H, J. There is no correlation between serotypes and PmpF genotypes, PmpF genotypes are age-related and have no relation to gender.