基于沙眼衣原体ompl基因限制性片段长度多态性和测序法用于女性生殖道感染沙眼衣原体基因分型

Chlamydia trachomais Genotyping Methods with ompl PCR-RFLP and Sequencing in Clinical Urogenital Samples of Women

沙眼衣原体D-K型见于女性生殖道感染,尤其是宫颈感染的主要病原体,基因分型针对编码主外膜蛋白(MOMP)编码基因(ompl)多态性,CT各型ompl基因长度略有差异,总长度均在1.1 kb左右,使用ompl基因限制性片段长度多态性(restriction fragment length polymorphism,RFLP)进行分型,但是不同的引物设计和实验条件,结果的判读不同,结合ompl基因测序法,能构建出不同实验条件下PCR-RFLP酶切图谱,便于临床判读,并且通过BLASTA及多序列比对发现ompl基因序列碱基变异。收集2010年1月至2014年5月在柳州市人民医院就诊的宫颈脱落细胞沙眼衣原体阳性女性,结合omp1基因测序法进行分型,建立反应体系,通过ompl基因测序结果比对,确定本实验条件的参考酶切图谱,用于临床标本的CT分型检测。基于ompl基因的PCR-RFLP技术是目前沙眼衣原体基因分型临床可运用的方法。

CT has been classified into15 basic serotypes based on MOMP which coded by ompl gene through monoclonal or polyclonal immunogenic antibodies, and genotype D-K mainly causes Urogenital infection. Now PCRRFLP based on the 1.1 kb ompl gene in length were used for clinical research, but different primer selection and experiment condition may led to different result. To build up the PCR reaction system and form reference RFLP restriction maps by combination CT ompl PCR-RFLP and ompl sequencing. The cervical swabs of positive CT-NAAT were collected for genotyping in Liuzhou people's hospital from Jan, 2010 to May, 2014. We employed ompl gene PCR-RFLP combination with ompl gene sequencing to establish PCR reaction system, ompl gene sequencing and BLAST, to form reference restriction enzyme maps under our experimental condition. We established gene ompl PCR-RFLP restriction endonuclease maps to facilitate the genotyping of clinical samples. Meanwhile, we discover several variations in genome sequences through BLASTA and multiple sequence alignment.