摘要
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 develop a PCR assay for the rapid and sensitive detection of staphylococcal enterotoxins A - E （SEA, SEE） gene. Methods According to the sequences of SEA and SEE gene reported from genebank, primers were designed and used for PCR. The length of amplicon was 327 bp. To evaluate the specificity of PCR assay, 2 strains of SEA, SEE and 14 strains of non - SEA - E were tested by PCR; 1 strains of SEE was 10 - fold serially diluted and amplified by PCR to verify detection limit. PCR results were judged by electrophoretic analysis and DNA sequence analysis. Results With 2 strains of SEA and SEE, the results of PCR assay were observed. Amplification was not observed when 5 strains of non- SEA - E were tested. The specificity of PCR products was confirmed by DNA sequence analysis. In addition, the detection limit of PCR assay was 120 cfu/ml of SEE. Conclusions The results indicate that PCR assay is a rapid, specific and sensitive detection method for SEA and SEE gene, and it is suitable for clinical diagnosis and food safety applications.
出处
《实用预防医学》
CAS
2009年第1期246-248,共3页
Practical Preventive Medicine
