摘要
根据大肠杆菌的β葡糖醛酸酶基因(uid)、沙门氏菌侵袭性抗原保守基因(invA)和金黄色葡萄球菌的耐热核酸酶基因(nuc),分别设计3对特异性引物,通过对单管多重PCR扩增的特异性、敏感性以及扩增条件进行优化.结果表明:3对引物分别能扩增出147bp、570bp和280bp的特异性条带,反应条件优化后,3种目的菌在104 CFU/mL时均可同时扩增出较清晰条带(大肠杆菌和金黄色葡萄球菌在103 CFU/ml浓度下仍然可见到条带),仅沙门氏菌的检测比单基因PCR低一个稀释度,人工模拟果汁污染试验结果稳定.该方法操作简单、快速、特异性强,灵敏度高,能够实现对大肠杆菌、沙门氏菌和金黄色葡萄球菌3种菌的快速诊断检测和生产过程监控.
Based on the analysis for the sequences of uid gene in E. coli, conservauve lnvaslve anugcn gene （invA） in Salmonella and nuc gene in S. aureus, three pairs of special primers were designed and the specificity and sensitivity of PCR were analyzed for single gene. Further, the multiple PCR method was developed after analyzing and optimizing reaction conditiorL The results showed that aiming to E. coli, Salmonella and Staph- ylococcus aureus, on the optimized conditions, the anticipated PCR products were 147 bp, 570 bp and 280 bp, re- spectively, and the specificity was high,and the minimum detection limit was 104 CFU/mL,whieh was a dilute de- gree lower than that in the sensitivity of single PCR. The results were stable in simulated examination and in mar- keting examination. A rapid, specific and sensitive multiplex PCR system had been established and it was valuable for diagnosis and monitoring for E. coli, Salmonella and S. aureus.
出处
《甘肃农业大学学报》
CAS
CSCD
北大核心
2012年第3期124-128,133,共6页
Journal of Gansu Agricultural University
基金
现代苹果产业技术体系Cars-28
关键词
浓缩苹果汁
大肠杆菌
沙门氏菌
金黄色葡萄球菌
多重PCR
concentrated apple juice
Escherichia coli
Salmonella
Staphylococcus aureus
multiplePCR assay
