【摘要】 目的:建立一种快速、准确、特异、定量检测金黄色葡萄球菌及其肠毒素A、D的方法并探讨肠毒素在食物中毒引起腹泻中的意义。方法:选择femB、SEA和SED基因分别作为金黄色葡萄球菌菌株、肠毒素A、肠毒素D的靶序列,设计合成引物和探针;收集食物中毒导致腹泻患者粪便标本487份,并对其进行检测分析。结果:采用荧光聚合酶链反应检测金黄色葡萄球菌和肠毒素A的灵敏度为1.0×102拷贝,而检测肠毒素D的灵敏度为1.0×103拷贝;487份粪便标本中检出金黄色葡萄球菌femB基因72例(14.8%),检出产肠毒素A菌株为11例(15.3%,11/72),产肠毒素D菌株为9例(12.5%,9/72),同时产肠毒素A、D菌株1例(1.4%,1/72)。结论:应用Taqm an探针实时荧光PCR技术能够快速、准确检测金黄色葡萄球菌及其肠毒素,适合于大样本筛查。更多还原

【Abstract】 Objective:To develop a real-time fluorescence PCR assay for detecting the genes of staphyloccal enterotoxins A and D rapidly,and explore the clinical significance of staphyloccal enterotoxins.Methods:[WTBZ]The genes of SEA and SED were selected as target gene of staphyloccal enterotoxins A and D,and femB gene as a specific genomic marker for S.aureus.The primers and TaqMan probes were designed and synthesized.We investigated 487 stool samples of foodborne illness patients by real-time fluorescence PCR.Results:The sensitivity of the assay for Staphylococcus aureus and enterotoxins A was 1.0×10~2 copies,but the sensitivity of enterotoxins D was 1.0×10~3 copies.Among 487 samples,68 S.aureus strains were found,but femB gene were detected in 72 samples,11 samples showed the production of enterotoxins A,9 were positive for enterotoxins D,only one was positive for enterotoxins A and D.Conclusion:The TaqMan PCR is a rapid and sensitive method to detect the staphyloccal enterotoxins A and D and it is suitable for screening large numbers of samples simultaneously.更多还原