马鞍山市金黄色葡萄球菌鉴定及SPA基因多态性分析

Identification of Staphylococcus aureus and SpA polymorphisms in Ma'anshan City

目的鉴定分离自马鞍山的金黄色葡萄球菌,探索使用金黄色葡萄球菌SPA基因对菌株进行多态性分析。方法采集马鞍山市食品、食物中毒和腹泻患者标本,按照GB/4789.10-2003和WS/T80-1996进行金黄色葡萄球菌的分离培养。对分离的菌株,用过氧化氢酶和血浆凝固酶试验后,用生化实验对菌株进行鉴定。提取鉴定为金黄色葡萄球菌的DNA作为模板,PCR扩增菌株的SPA蛋白X区域并测序,测序结果提交数据库(http://www.seq-net.org/)进行分型,采用分型软件Ridom Staph Type对分型结果进行聚类分析。结果 共分离和鉴定54株金黄色葡萄球菌,其中食品、食物中毒和腹泻标本分别为36株、11株和7株。所有菌株的过氧化氢酶及血浆凝固酶均为阳性,生化实验鉴定为金黄色葡萄球菌。54株菌株可分成19个型,分别为t 189型11株、t 701型9株、t 091型7株、t 1376型5株、t 011型、t 127型各2株、t 084型、t 163型、t 459型、t 547型、t 548型、t 791型、t 954型、t 1544型、t 4336型、t 5269型、t 5353型各1株,同时发现2个新型,分别为t 5269和t 5353型。两起食物中毒的型别分别为t 189型和t 701型。结论 SPA基因分型方法具有快速、易于标准化的特点,可用于食物中毒的流行病溯源。

Objective To identify Staphylococcus aureus isolated from Ma＇anshan city,and explore the analysis of SpA gene polymorphism of the strains.Methods The samples from food,food poisoning and diarrhea cases in Ma＇anshan city were collected to isolate and culture S.aureus according to GB/4789.10-2003 and WS/T80-1996.The strains were tested with catalase and coagulase tests,biochemical experiments were used to identify the strain.S.aureus DNA were extracted as template,PCR amplification of the X region of SpA protein from isolates and sequencing,the sequencing results submit to the database（http：//www.seqnet.org/） for typing,cluster analysis was used with the Ridom Staph software.Results A total of 54 S.aureus were isolated and identified,including food,food poisoning and diarrhea specimens,36,11 and 7 strains respectively.All strains were positive of hydrogen peroxide enzymes and coagulase tests,and biochemical experiments identified as S.aureus.54 strains can be divided into 19 types,including type t189（11 strains）,t701（9）,t091（7）,t1376（5）,t011（2）,t127（2）,each type of t084,t163,t459,t547,t548,t791,t954,t154,t4336,t5269 and t5353 with one strain.Furthermore,2 new types（t5269 and t5353）were found.The types of 2 food poisonings were t189 and t701.Conclusion With the advantage of rapid and easy standardization,SpA genotyping method can be used for tracing epidemic source of food poisoning.