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目的建立一种用PCR快速敏感地检测金黄色葡萄球菌肠毒素A、E的方法。方法根据genebank报道的编码金黄色葡萄球菌肠毒素A、E的基因序列,设计一对特异性引物来扩增靶基因片段,长度为327 bp,通过对金黄色葡萄球菌产肠毒素A、E菌株和14株非金黄色葡萄球菌产肠毒素A、E菌株进行检测,评价该方法的特异性;对金黄色葡萄球菌产肠毒素E菌株做系列10倍稀释,进行PCR检测,对其敏感性进行分析;PCR扩增产物进行电泳和测序鉴定。结果金黄色葡萄球菌肠毒素A、E菌株出现PCR扩增反应,14株非金黄色葡萄球菌产肠毒素A、E菌株没有出现PCR扩增反应,通过测序分析证实了PCR产物的特异性;PCR检测金黄色葡萄球菌肠毒素E基因的检测下限为120 cfu/ml。结论建立了一个快速、特异、敏感的金黄色葡萄球菌肠毒素A、E基因的PCR检测方法,可用于金黄色葡萄球菌肠毒素的临床诊断和食品安全监测。
Objective To establish a rapid and sensitive method for detecting Staphylococcus aureus enterotoxin A and E by PCR. Methods A pair of specific primers was designed to amplify the target gene fragment according to the gene sequence of the gene encoding Staphylococcus aureus enterotoxins A and E reported by genebank. The length of the target gene fragment was 327 bp. Staphylococcus aureus enterotoxins producing strains A and E And Staphylococcus aureus Staphylococcus aureus toxin A, E strains were tested to evaluate the specificity of the method; Staphylococcus aureus Enterotoxin E strain to do a series of 10-fold dilution, PCR detection, its sensitivity Analysis; PCR amplification products were electrophoresis and sequencing. Results Staphylococcus aureus enterotoxin A and strain E showed PCR amplification reaction. No staphylococcal enterotoxins producing strains A and E were detected in 14 strains of Staphylococcus aureus. The PCR products were confirmed by sequencing analysis. PCR Detection of Staphylococcus aureus enterotoxin E gene detection limit of 120 cfu / ml. Conclusion A rapid, specific and sensitive PCR detection method of Staphylococcus aureus enterotoxin A and E genes was established and could be used for clinical diagnosis and food safety monitoring of Staphylococcus aureus enterotoxin.