摘要
目的建立一种快速鉴定携带耐药基因和肠毒素A基因的金黄色葡萄球菌的方法。方法根据金黄色葡萄球菌耐热核酸酶基因nuc、甲氧西林耐药基因mecA和肠毒素A基因sea的序列,设计特异性的引物和不同荧光素标记的TaqMan探针,通过反应体系和反应条件的优化,建立了能同时检测鉴定耐甲氧西林和产肠毒素A的金黄色葡萄球菌的多重荧光PCR方法。结果该方法特异性好,可在同一个PCR反应体系中同时检测金黄色葡萄球菌的3种基因成分,检测效率高。灵敏度可以达到2×10-7μg/μl。结论该方法可用于食品样品的检测和流行病学分离株的快速鉴定。
Objective To establish a multiplex fluorescent real-time PCR assay for the detection of methicillin-resistance gene and enterotoxin gene A of Staphylococcus aureus.Methods We designed the specific primers and TaqMan probes targeting sea,mecA and nuc gene of Staphylococcus aureus.We used different fluorescents to label the probes.After optimization of reaction condition,we set up a multiplex fluorescent real-time PCR assay to detect Staphylococcus aureus rapidly and accurately.Results The result demonstrated that this assay was specific,which can identify three kinds of genes from Staphylococcus aureus in one reaction.The lowest detection limit was 2×10-7μg/μl.Conclusion This assay can be used in food sample detection and rapid identification of Staphylococcus aureus.
出处
《疾病监测》
CAS
2011年第12期999-1003,共5页
Disease Surveillance
基金
科技支撑计划项目(No.2009BAD9B06
2011BAD09B02
2011AA100902)~~
