泌尿生殖道标本中沙眼衣原体的直接检测及基因分型

Direct Detection and Genotyping of Chlamydia trachomatis in Urogenital Specimens

应用聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析，建立了沙眼衣原体（Ct）直接检测及基因分型的方法。与细胞培养法比较，评价其检测的敏感性。400份性病门诊患者的泌尿生殖道标本同时用培养和质粒PCR检测Ct，结果29份标本培养阳性，其中28份标本质粒PCR阳性，另有5份培养阴性的标本质粒PCR阳性。对培养和质粒PCR阳性的标本进一步做主要外膜蛋白基因（ompl）PCR或ompl巢式PCR，结果32份标本omplPCR或ompl巢式PCR阳性。将阳性标本的ompl产物用AluI和MspI等限制性内切酶酶切，产生的酶切带谱与标准株带谱比较，以鉴定阳性标本的基因型。结果E型为13份标本，F：6、G：4、D：3、J：2、K：2、H：1、非典型：1。研究表明：PCR－RFLP可直接检测及基因分型泌尿生殖道标本中的Ct，为一种敏感的血清分型替代方法。

A method was developed to detect and genotype Chlarnydia trachomatis by using polymerase chain reaction (PCR) and restriction fragment length polymorphisrn(RFLP). The sensitivity of this method was evaluated by comparing with cell culture. A total of 400 urogenital specimens collected frorn patients attending sexually transmitted disease clinic were first screened for C. trachomatis by both cell culture and plasmid PCR. Twenty-nine specimehs were culture positive,of which 28 were positive by plasmid PCR. In addition. of the .culture negative group,five samples were founded positive by plasmid PCR. Subsequently,positive samples by plasmid PCR and culture were tested by PCR against the major outer memberane protein gene(ompl) or by ompl nested PCR. The ompl PCR products were digested by restriction endonuclease AluI and MspI. The restriction pattern of positive specimens were compared with that of reference strains to determine the genotypes of C. trachomatis. Analysis of 32 C.trachomatis positive specmens gave the following results: 13 E strains, 6F,4G, 3D, 2J, 2K, 1H and 1 atypical genotype. The results showed that direct PCR-based RFLP analysis were feasible for detection and genotyping of C.trachomatis in Urogenital Specimens and a good Qlternative to Serotyping method for differentiation of C.trachomatis.