CAJ | 학술논문

目的：建立快速检测食源性耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus, MRSA)Taqman探针双色荧光PCR方法。方法根据金黄色葡萄球菌种属鉴定nuc基因和MRSA决定因子mecA基因,设计合成引物探针,建立双色荧光PCR扩增体系。利用所建立的方法检测特异性及灵敏度。将金黄色葡萄球菌依次传代培养,检测不同代次的菌株验证方法的稳定性,并对实际样品分离株进行检测验证方法的可行性与实用性。结果该方法可准确并特异性检测出 MRSA 和甲氧西林敏感金黄色葡萄球菌(methicillin-susceptible Staphylococcus aureus, MSSA),检测 MRSA 的 nuc 基因和 mecA 基因的灵敏度可达2.7×103 CFU/mL,不同代次的菌株的检测结果一致。结论本实验所建立的双色荧光PCR检测方法具有良好的特异性、灵敏度及稳定性,可用于快速检测食源性MRSA。
목적：건립쾌속검측식원성내갑양서림금황색포도구균(methicillin-resistant Staphylococcus aureus, MRSA)Taqman탐침쌍색형광PCR방법。방법근거금황색포도구균충속감정nuc기인화MRSA결정인자mecA기인,설계합성인물탐침,건립쌍색형광PCR확증체계。이용소건립적방법검측특이성급령민도。장금황색포도구균의차전대배양,검측불동대차적균주험증방법적은정성,병대실제양품분리주진행검측험증방법적가행성여실용성。결과해방법가준학병특이성검측출 MRSA 화갑양서림민감금황색포도구균(methicillin-susceptible Staphylococcus aureus, MSSA),검측 MRSA 적 nuc 기인화 mecA 기인적령민도가체2.7×103 CFU/mL,불동대차적균주적검측결과일치。결론본실험소건립적쌍색형광PCR검측방법구유량호적특이성、령민도급은정성,가용우쾌속검측식원성MRSA。
Objective To establish a double-colored real-time fluorescence PCR method of detection of foodborne Staphylococcus aureus by Taqman probe. Methods Two pairs of specific primers and Taqman fluorescent probe were designed and synthesized according to nuc gene for species identification of S.aureus and mecA gene for determination gene of MRSA. Double-colored real-time fluorescence PCR amplification system was developed. The specificity and sensitivity were tested by the established method. In order to validate the stability of method, strains of different generations were tested, after strains of S.aureus were subcultured successively. The feasibility and practicality were studied by detecting isolates from practical samples. Results MRSA and MSSA could be detected accurately, rapidly and specifically in this study. The sensitivity of detection method was 2.7×103 CFU/mL for nuc and mecA gene of MRSA. The detection results of different generations were consistent. Conclusion This method has high specificity, sensitivity and stability for double-colored real-time fluorescence PCR method, and it can be applied to rapid detection and identification of foodborne MRSA.