淋球菌opa基因分型方法的建立与评估

Opa-typing of Neisseria gonorrhoeae： establishment and evaluation of a new method

目的探讨建立淋球菌opa基因分型方法，并对建立的实验室方法进行流行病学资料验证。方法采用PCR—RFLP建立淋球菌opa基因分型方法，opa基因多态性分析使用特定的Gelcompar软件。选取淋病患者26例，其中有三对患者两两之间具有明确的性关系，而其他患者之间无明确性接触史。研究用收集到的已知流行病学资料验证opa基因分型方法的可靠性。结果建立PCR—RFLP为基础的opa基因分型方法。26株淋球菌opa基因经过两种限制性内切酶（TaqⅠ与HpaⅡ）分别酶切后，发现两两之间存在明确性联系的3对菌株其opa基因被酶切后具有相同（100％）的长度多态性特性，而无明确性联系的菌株其opa基因被酶切后未发现具有类似相同的长度多态性。结论实验室结果与患者所提供的性接触信息完全吻合。opa基因分型方法是一种较可靠的性网络调查工具。

Objective To establish a molecular typing method （opa-typing） for Neisseria gonorrhoeae, and to evaluate its performance based on epidemiological data. Methods Twenty-six gonorrhea patients were recruited from March to April 2006 at two sites, including 17 cases from the STD Clinic of Institute of Derma- tology, Chinese Academy of Medical Sciences and Peking Union Medical College and 9 cases from the STD Clinic of Shandong Provincial Institute of Dermatology. Of the 26 patients, 6 were from three known sexual links, while the remaining 20 patients did not have any sexual contact with each other. The opa gene was amplified by using PCR from Neissen＇a gonorrhoeae isolates from these patients followed by overnight digestion with restriction enzymes Taq ｜ and Hpa lI. The enzyme digestion band patterns were analyzed using the Gel- Compar program. Results The opa gene was successfully amplified from all the 26 Neisseria gonorrhoeae strains, and restrictedly digested by endonucleases Taq ｜ and Hpa ]]. Identical band patterns were observed between patients with sexual links, but not among the remaining 20 patients. Conclusions The results of opatyping of Neisseria gonorrhoeae coincide with the information on sexual behaviour provided by patients. Opatyping may serve as a reliable tool in sexual network analysis.