基于上转发光技术的奶粉及牛奶中黄曲霉毒素M1快速定量检测方法研究  

Rapid detection of aflatoxin M1 in milk powder and milk based on up-converting phosphor technology

作　　者：刘晓[1] 王立平[2] 周蕾[3] 林承喜 杨瑞馥[3] 邱景富[1] 

机构地区：[1]重庆医科大学公共卫生与管理学院,重庆400016 [2]国家食品质量安全监督检验中心,北京100094 [3]军事医学科学院微生物流行病研究所,北京100071 [4]中海智(北京)科技有限公司,北京100027

出　　处：《军事医学》2014年第11期850-854,共5页Military Medical Sciences

基　　金：国家科技重大专项资助项目(2011ZX10004-001;2012ZX10004801-002-004;2012ZX10004801-004-015);国家863项目(2013AA032205)

摘　　要：目的利用上转发光免疫层析技术（AFM1-UPT-LF）建立奶粉及牛奶中黄曲霉毒素M1的快速检测方法。方法以UCP颗粒作为标记物,无需样品前处理,采用竞争免疫反应建立AFM1-UPT-LF方法。以AFM1阴性的奶粉、牛奶配制系列浓度AFM1标准品,确定检测方法的灵敏度、特异性、标准曲线与定量范围、精密度、准确度及回收率,并对实际样品进行定性和定量检测评价。结果利用AFM1-UPT-LF可实现奶粉及牛奶中AFM1定性及定量检测,20 min内即可完成。奶粉中检测下限可达0.1μg/kg,在0.1-0.7μg/kg间具有较好线性（r=-0.99359,P〈0.01）,牛奶中检测限为0.3μg/L,在0.3-0.7μg/L间具有较好线性（r=-0.99938,P〈0.05）。在定性检测中,对奶粉检测灵敏度为100%,特异性为85%,受试者工作特征（ROC）的曲线下面积（AUC）为0.978±0.036;牛奶检测中灵敏度为81%,特异性为88%,ROC的AUC为0.891±0.099;与LC-MS结果进行2检验,P值均大于0.05,两种方法差异无显著性意义。在定量检测评价中,AFM1-UPT-LF对于奶粉及牛奶中AFM1检测均具有较好回收率,与LC-MS比较差异无显著性意义（奶粉：t=0.66,P〉0.05;牛奶：t=1.01,P〉0.05）。结论利用UPT-LF所建立的AFM1-UPT-LF方法可实现奶粉及牛奶中AFM1定性及定量快速检测,检出限满足国标限量标准要求;该法无需样品处理,操作简便,可满足食品安全中对现场快速检测的需求。Objective To develop and evaluate an up-converting phosphor technology-based lateral flow assay （UPT- LF） for detection of aflatoxin M1 （AFM1） in milk powder and milk. Methods AFM1-UPT-LF was established with upconverting phosphor （UCP） nano-particles as the bio-label of competitive mode based LF assay. Sensitivity, quantitative ability and precision were evaluated using simulated AFMl-postive samples with serial standard concentrations. The qualitative and quantitative detection performance of AFM1-UPT-LF was evaluated with reference to liquid chromatography-mass spectrography （LC-MS） to detect samples of milk powder and milk simultaneously. Results AFM1-UPT-LF could conduct qualitative and quantitative detection without sample pretreatment within 20 min. The detection limit of AFM1-UFF-1,F reached 0. 1 μg/kg in milk powder and 0.3 μg/L in milk. There was good linearity ranging from 0.1 to 0.7μg/kg and 0.3 to 0.7 μg/L for milk powder and milk, respectively. The sensitivity, specificity and receiver operating characteristic （ROC） area under the curve（AUC） of AFM1-UPT-LF for qualitative result could meet the need of national standards fnr AFMl limit in dairy products. After statistical analysis, there was no significant difference （milk powder： t = 0. 66, P 〉 0.05 ; milk ： t = 1.01, P 〉 0.05） between AFM1-UPT-LF and LC-MS for quantitative detection. Conclusion The good qualitative and quantitative detection performance of AFM1-UPT-LF for milk powder and milk makes possible on-site rapid detection of AFM1 in dairy products quantitatively.

关 键 词：上转发光免疫层析技术 侧流术 黄曲霉毒素M1 快速检测 定量检测 奶制品 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]