摘要
目的建立快速检测葡萄球菌肠毒素C基因（staphylococcal enterotoxin C，SEC）PCR方法。方法根据SEC基因的序列，设计特异性PCR引物，建立扩增体系，琼脂糖凝胶电泳检测扩增的靶基因片段。通过分析产SEC菌株和对照菌株PCR产物，评价方法的特异性；通过对产SEC金黄色葡萄球菌定量样本系列稀释后进行PCR检测，分析方法的敏感性；并运用该方法分析实验室既往检出的30株金黄色葡萄球菌SEC基因的携带情况。结果建立了金黄色葡萄球菌SEC基因PCR快速检测方法，该法PCR扩增产物长度为490bp，未见假阳性结果，特异度良好；灵敏度较高，反应体系中有32cfu的产SEC金黄色葡萄球菌即可检出；30株本地分离的金黄色葡萄球菌SEC基因携带率20％。结论PCR法可以快速、敏感地检测SEC基因，是金黄色葡萄球菌食物中毒诊断可以利用的有效工具。
Objective To establish a polymerase chain reaction（PCR） method for rapid and sensitive detection of Staphylococcal enterotoxin C （SEC） gene. Methods According to the SEC gene sequences,to design specific PCR primers to create amplification system, the amplified target gene fragment were tested by agarose gel electrophoresis. Specificity of this method were evaluated by analyzing the production the SEC strain and the control strain PCR product. Sensitivity were analyzed by series dilution the SEC positive Staphylococcus quantitative sample. SEC gene carryingrate of 30 Staphylococcus aureus preserved in the laboratory were analysised by the method. Results A rapid PCR method was established for the detection SEC of Staphylococcus,a product length 490 bp could be observed from positive sample,and no false-positive results were fined verify the specificity and higher sensitivity. There were 32cfu SEC positve Staphylococcus aureus that couldan be detected in the reaction system. The carring rate of SEC gene of 30 Staphylococcus aureus was 20%. Conclusion The PCR method can be used in detection for the SEC gene of Staphylococcus, and was useful tool for the rapid and sensitive diagnosis of Staphylococcus food poisoning.
出处
《现代检验医学杂志》
CAS
2013年第2期57-59,62,共4页
Journal of Modern Laboratory Medicine
