应用核酸层析技术快速检测奶粉中阪崎肠杆菌的研究  

Study on nucleic acid lateral flow method for rapid detection of Enterobacter sakazakii from powdered milk

作　　者：王丽丽[1] 杨海荣[1] 赵瑜[1] 何艳玲[1] 

机构地区：[1]中国检验检疫科学研究院,北京100123

出　　处：《中国乳品工业》2013年第6期39-42,共4页China Dairy Industry

基　　金：国家质量监督检验检疫总局项目(2011IK250)

摘　　要：结合PCP与胶体金免疫层析试纸条技术建立了核酸层析检测技术用于奶粉中阪崎肠杆菌的快速检测。该技术根据寡-1,6-葡萄糖苷酶基因设计引物,并分别标记生物素和地高辛,通过试纸条上肉眼可见的红色条带对扩增结果进行判断。通过11株阪崎肠杆菌及27株常见致病菌的检测结果说明其特异性强,阪崎肠杆菌纯菌培养检测灵敏度达到103mL-1,人工污染样品检测限达到0.08 g-1,检测奶粉样品时与传统方法(GB/T 4789.40-2010)相比,无显著性差异(x2=0,P>0.05)。该方法灵敏度高、快速、特异性强,可以很好地应用于奶粉以及其他食品中阪崎肠杆菌的检测与鉴定。This article presents a new Nucleic Acid Lateral Flow （NALF） method, based on PCR. with Immune chromatographic test strip, for the rapid detection of E. sakazakii from powdered milk. NALF is based on oligo-1, 6-ghicosidase gene, with two specific primers, of which one was labelled with biotin and the other with digoxigenin, the pcr result can be interpretated directly according to the visible lines on a matched test strip. Specificity of NALF is tested against two-type strains, nine isolates of E.sakazakii and other twenty seven non-E, sakazakii species.The detection sensitivity is 103 mL-1 in pure culture, detection limit of artificially contaminated samples is 0.08 g-1 powdered milk after enrichment. And there is no observed difference （xa=0, P〉0.05） when compared with classical microbiological detection method （GB/T 4789. 40-2010）. This article demonstrates that NALF is easy; rapid, efficient and may has applications for the detection of E. sakazakii from pow- dered milk or other foods.

关 键 词：阪崎肠杆菌 聚合酶链式反应 核酸层析 胶体金 奶粉 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]