PCR法检测葡萄球菌肠毒素A基因

Detection of gene encoding Staphylococcal enterotoxin A by PCR

目的建立PCR方法快速检测葡萄球菌肠毒素A(Staphylococcal enterotoxin A,SEA)基因。方法根据SEA基因的序列,设计PCR引物特异性扩增靶基因片段,对1株产SEA金黄色葡萄球菌和9株对照菌株进行PCR检测,评价其特异性和敏感性。另检测30株金黄色葡萄球菌SEA基因的携带情况。结果建立PCR方法快速检测SEA基因,扩增产物长度为439bp。PCR反应体系中有29cfu的产SEA金黄色葡萄球菌即可检出SEA基因,30株分离的金黄色葡萄球菌中有10株携带有SEA基因。结论 PCR法可以快速、敏感地检测SEA基因,为金黄色葡萄球菌食物中毒诊断提供依据。

Objective To establish a polymerase chain reaction（PCR） assay for the rapid and sensitive detection of Staphylococcal enterotoxin A （SEA） gene. Methods According to the sequences of the SEA gene, primers were selected and were used to amplify SEA gene. The specificity of the assay was determined by detection of 1 SEA positive Staphylococcus aureus strain and 9 non-Staphylococcus aureus strains using PCR. The detection limit of the assay for SEA positive Staphylococcus aureus strain was 10-fold serially diluted and was amplified by PCR. The SEA gene in 30 Staphylococcus aureus strains were detected by PCR. Results The PCR assay for the rapid and sensitive detection of SEA gene was well established. The length of amplification was 439bp. The detection limit for SEA gene was 29 CFU. Among 30 Staphylococcus aureus strains 10 strains were positive for SEA gene. Conclusion The PCR assay can rapidly and exactly detect the SEA gene, and can provide evidence for the rapid diagnosis of food poisonings due to Staphylococcus aureus infection..