摘要
目的:了解食源性金黄色葡萄球菌中耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)及对氨基糖苷类庆大霉素、卡那霉素抗生素的耐药性情况,并建立快速检测鉴定金黄色葡萄球菌耐药性多重PCR方法。方法:对金黄色葡萄球菌进行增菌培养,并提取DNA作为模板,以nuc基因作为菌属鉴定基因,mec A基因作为MRSA检测基因,aac A-aph D及aph(3')-Ⅲa基因作为耐氨基糖苷类庆大霉素、卡那霉素的检测目的基因,建立四重PCR检测方法并进行特异性及灵敏度评价;利用所建立的方法检测72株食品及食物中毒患者来源的金黄色葡萄球菌。结果:成功建立四重PCR检测方法,且72株金黄色葡萄球菌均有nuc基因检出,mec A基因与MRSA检测符合率达100%,aac A-aph D、aph(3')-Ⅲa基因与庆大霉素、卡那霉素耐药菌株的符合率达95.12%。结论:本实验所建立的多重PCR方法灵敏度高、特异性好,可以快速检测出MRSA,并对氨基糖苷类庆大霉素、卡那霉素耐药菌株检测具有较高的准确率。
Objective: To investigate the prevalence of methicillin-resistant Staphylococcus aureus MRSA and resistance to Aminoglycoside gentamicin, kanamycin in foodborne S.aureus,and establish a rapid quadruple-PCR method for detection and identification antibiotic resistance of S.aureus.Methods: Genomic DNA was extracted as template for PCR from S.aureus recoveried and cultured in enrichment medium.According to nuc for Genus,mecA for MRSA, aacA-aphD and aph (3') - Ⅲ a for Gentamicin and kanamycin, quadruple- RCR detection method was developed, its sensitivity and specificity were evaluated; a total of 72 strains of S.aureus from food and patients with food poisoning were detected by quadruple RCR method. Results. Quadruple- RCR detection method was established.?2 strains of S.aureus all had nuc,the coincidence rate of mecA and MRSA was 100% ,the coincidence rate of aacA-aphD, aph (3') - Ⅲ a and gentamicin, kanamycin was 95.12%.Conclusion : this established quadruple -RCR method was specific, sensitive. MRSA could be tested rapidly, and there was high accurate rate for detection of gentamicin,kanamycin resistant strains of S.aureus.
出处
《食品工业科技》
CAS
CSCD
北大核心
2015年第1期303-308,共6页
Science and Technology of Food Industry
基金
科技部质检公益性专项课题(201210043)