CAJ | 학술논문

目的：建立一种快速、简便的模板提取方法用于 PCR 检测粪便中的金黄色葡萄球菌。方法利用 FTA滤膜，无需增菌，直接从人工污染金黄色葡萄球菌的粪便中提取 DNA 模板，以金黄色葡萄球菌耐热核酸酶基因（nuc）为靶基因，经过 PCR 扩增得到279 bp 的产物。通过凝胶电泳，和凝胶成像技术观察结果。结果使用 FTA 滤膜可有效的提取 DNA 模板，消除了样本中的 PCR 抑制因子，通过 PCR 方法检测人工污染的粪便中的金黄色葡萄球菌，检出限为102 cfu/ml，可在6 h 内完成检测。结论FTA 滤膜用于 PCR 检测粪便中的金黄色葡萄球菌操作简便、快速。使用 FTA 滤膜法制备模板 DNA，为食源性致病菌快速检测构建了一个技术平台。
목적：건립일충쾌속、간편적모판제취방법용우 PCR 검측분편중적금황색포도구균。방법이용 FTA려막，무수증균，직접종인공오염금황색포도구균적분편중제취 DNA 모판，이금황색포도구균내열핵산매기인（nuc）위파기인，경과 PCR 확증득도279 bp 적산물。통과응효전영，화응효성상기술관찰결과。결과사용 FTA 려막가유효적제취 DNA 모판，소제료양본중적 PCR 억제인자，통과 PCR 방법검측인공오염적분편중적금황색포도구균，검출한위102 cfu/ml，가재6 h 내완성검측。결론FTA 려막용우 PCR 검측분편중적금황색포도구균조작간편、쾌속。사용 FTA 려막법제비모판 DNA，위식원성치병균쾌속검측구건료일개기술평태。
Objective To establish an new template preparation method for rapid and sensitive PCR detection of Staphylococcus aureus in feces .Methods FTA filter membrane was used to extract DNA in Staphylococcus aureus from artificially contaminated feces.Primers targeting the thermostable nuclease gene (nuc) were used to amplify a 279 bp DNA fragment which was confirmed by DNA sequencing .Results Application of FTA filter membrane could effectively extract DNA template and removed PCR inhibitive factor in specimens .In the detection of Staphylococcus aureus from artificially contaminated feces by PCR,the detection limit was 102cfu/ml,which could be finished within 6 hours.Conclusion PCR amplification using FTA filter membrane provides a faster and more sensitive method of detecting Staphylococcus aureus in feces.At the same time,which establishes a technological platform for rapid detection of foodborne pathogenic bacteria .