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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 and to explore the use of Staphylococcus aureus SPA gene polymorphism analysis of the strains. Methods The samples of food, food poisoning and diarrhea in Ma’anshan City were collected and the Staphylococcus aureus was isolated and cultured according to GB / 4789.10-2003 and WS / T80-1996. Isolates were isolated and tested for catabolism with catalase and plasma coagulase. The DNA identified as Staphylococcus aureus was extracted as a template and the SPA protein X region of the strain was amplified by PCR and sequenced. The sequencing results were submitted to the database (http://www.seq-net.org/) for typing, and typing software Ridom Staph Type cluster analysis of the classification results. Results A total of 54 strains of Staphylococcus aureus were isolated and identified, of which 36, 11 and 7 were food, food poisoning and diarrhea respectively. All strains of catalase and plasma coagulase were positive, biochemical experiments identified as Staphylococcus aureus. The 54 strains were divided into 19 types, including 11 strains of t 189, 9 strains of t 701, 7 strains of t 091, 5 strains of t 1376, 2 strains of t 011 and 2 strains of t 127, t 084, t 163 type, t 459 type, t 547 type, t 548 type, t 791 type, t 954 type, t 1544 type, t 4336 type, t 5269 type and t 5353 type respectively. Two novel types For the t 5269 and t 5353 type. Two types of food poisoning were t 189 and t 701 type. Conclusion The method of SPA genotyping is rapid and easy to standardize and can be used to trace the epidemic of food poisoning.