摘要
目的:研究阪崎肠杆菌的环介导等温扩增(loop-mediated isothermal amplification,简称LAMP)方法,实现对食品中阪崎肠杆菌的快速检测。方法:针对阪崎肠杆菌16S rRNA基因(16S rDNA)及外膜蛋白A(OmpA)基因设计LAMP引物,扩增后通过电泳或加入显色剂SYBR-GREENⅠ检测。结果:LAMP法能有效特异地检测出阪崎肠杆菌;以16S rRNA引物检测阪崎肠杆菌的最低检出限为32CFU/mL,用OmpA引物检测阪崎肠杆菌的最低检出限达3CFU/mL,扩增产物加入SYBR-GREENⅠ在紫外条件下可见荧光,与电泳检测具有同样的灵敏度。结论:LAMP法可实现对阪崎肠杆菌的快速检测,在食品检测中具有广阔的应用前景。
Objective: To develop a loop-mediated isothermal amplification (LAMP) method for rapid detection of Enterobacter sakazakii in food. Methods: LAMP primers were designed on the basis of 16S rRNA gene (16S rDNA) and outer membrane protein A gene (OmpA) in Enterobacter sakazakii. The LAMP products were evaluated by electrophoresis or SYBR-GREEN L Results: The detection limit of Enterobacter sakazakii by LAMP assay was 32 CFU/mL using 16S rRNA primers or 3 CFU/mL using OmpA primers. The similar detection sensitivity of LAMP products using SYBR-GREEN I and electrophoresis was observed. Conclusion: LAMP method was effective and sensitive for rapidly detecting Enterobacter sakazakii, which will have a broad application prospect in food.
出处
《食品科学》
CAS
CSCD
北大核心
2010年第22期322-325,共4页
Food Science
基金
武汉市科技局对外科技合作与交流计划项目(201070934341)
武汉工业学院研究生创新基金重点项目(09cx002)
关键词
阪崎肠杆菌
环介导等温扩增
快速检测
Enterobacter sakazakii
loop-mediated isothermal amplification
detection