免疫磁分离和可视化金纳米杂交探针法快速检测婴幼儿配方乳粉中的阪崎克罗诺杆菌  

Rapid Detection of Cronobacter sakazakii in Powdered Infant Formula by Immunomagnetic Separation Coupled with Visual Nanogold Hybridization Probe

作　　者：姜毓君[1] 杨涛 满朝新[1] 周文琦[1] 王蕊[1] 陈思涵 杨鑫焱 付世骞 高平娉 任玉伟 张微[1] JIANG Yujun;YANG Tao;MAN Chaoxin;ZHOU Wenqi;WANG Rui;CHEN Sihan;YANG Xinyan;FU Shiqian;GAO Pingping;REN Yuwei;ZHANG Wei(Key Lab of Dairy Science,Ministry of Education,College of Food Science,Northeast Agricultural University,Harbin 150030,China)

机构地区：[1]东北农业大学食品学院,乳品科学教育部重点实验室,黑龙江哈尔滨150030

出　　处：《食品科学》2021年第16期224-232,共9页Food Science

基　　金：国家自然科学基金面上项目(31871828)。

摘　　要：利用免疫磁分离和金纳米杂交探针策略,开发一种快速和灵敏的可视化检测婴幼儿配方乳粉(powdered infant formula,PIF)中阪崎克罗诺杆菌(Cronobacter sakazakii)方法。抗体功能化的磁颗粒捕获C.sakazakii,金纳米探针对聚合酶链式反应的产物进行分析,这种探针法可以替代传统的电泳。免疫磁颗粒是由100μL磁颗粒、120μL碳化二亚胺盐酸盐、80μL C.sakazakii单克隆抗体和体积分数0.02%的吐温-20制备。特异性检测中仅有C.sakazakii是阳性结果。在纯培养基和不经预富集的人工污染PIF中,分别能检测到10^(2) CFU/mL和10^(3 )CFU/g的C.sakazakii。经过3 h的预富集,此方法可以在PIF中检测到低至4.5×10^(1) CFU/g的C.sakazakii。可视化检测、电泳和紫外扫描光谱结果一致。因此,免疫磁分离和金纳米探针的方法可以替代凝胶电泳,对于快速检测C.sakazakii具有重要的意义。此方法将有助于检测PIF样本的C.sakazakii。A rapid and sensitive assay for detecting Cronobacter sakazakii in powdered infant formula(PIF)was developed by utilizing immunomagnetic separation(IMS)combined with visual gold nanoparticle(GNP)hybridization probe.C.sakazakii was captured via antibody-functionalized magnetic particles(MPs)and the polymerase chain reaction amplicon of the target bacteria was analyzed by GNP instead of traditionally used electrophoresis.Immunomagnetic particles(IMPs)were obtained by mixing together 100μL of MPs,120μL of carbodiimide hydrochloride(EDC)and 80μL of monoclonal antibody against C.sakazakii with 0.02%(V/V)added Tween-20.Specificity test showed positive results only for C.sakazakii.C.sakazakii could be detected at 10^(2)CFU/mL and 10^(3)CFU/g in pure culture and artificially contaminated PIF without pre-enrichment,respectively.After 3 h of pre-enrichment,this assay was able to detect as low as 4.5×10^(1)CFU/g of C.sakazakii in PIF.The results of the visual assay were consistent with those of electrophoresis and ultraviolet scanning spectroscopy.Therefore,the IMS-GNP probe assay proved to be an alternative to gel electrophoresis for the rapid detection of C.sakazakii in PIF samples.

关 键 词：阪崎克罗诺杆菌 免疫磁分离 金纳米颗粒 婴幼儿配方乳粉 可视化检测 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]