CAJ | 학술논문

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χ2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。
목적：이용상전발광면역층석기술（ AFM1-UPT-LF）건립내분급우내중황곡매독소M1적쾌속검측방법。방법이UCP과립작위표기물，무수양품전처리，채용경쟁면역반응건립AFM1-UPT-LF방법。이AFM1음성적내분、우내배제계렬농도AFM1표준품，학정검측방법적령민도、특이성、표준곡선여정량범위、정밀도、준학도급회수솔，병대실제양품진행정성화정량검측평개。결과이용AFM1-UPT-LF가실현내분급우내중AFM1정성급정량검측，20 min내즉가완성。내분중검측하한가체0．1μg／kg，재0．1～0．7μg／kg간구유교호선성（r＝－0．99359， P＜0．01），우내중검측한위0．3μg／L，재0．3～0．7μg／L간구유교호선성（r＝－0．99938，P＜0．05）。재정성검측중，대내분검측령민도위100％，특이성위85％，수시자공작특정（ROC）적곡선하면적（AUC）위0．978±0．036；우내검측중령민도위81％，특이성위88％， ROC적AUC위0．891±0．099；여LC-MS결과진행χ2검험，P치균대우0．05，량충방법차이무현저성의의。재정량검측평개중，AFM1-UPT-LF대우내분급우내중AFM1검측균구유교호회수솔，여LC-MS비교차이무현저성의의（내분：t＝0．66，P＞0．05；우내：t＝1．01，P＞0．05）。결론이용UPT-LF소건립적AFM1-UPT-LF방법가실현내분급우내중AFM1정성급정량쾌속검측，검출한만족국표한량표준요구；해법무수양품처리，조작간편，가만족식품안전중대현장쾌속검측적수구。
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 up-converting phosphor ( UCP) nano-particles as the bio-label of competitive mode based LF assay .Sensitivity, quantitative ability and precision were evaluated using simulated AFM 1-postive samples with serial standard concentrations .The qualita-tive and quantitative detection performance of AFM 1-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-UPT-LF reached 0.1 μg/kg in milk powder and 0.3 μg/L in milk.There was good linearity ranging from 0.1 to 0.7 μg/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 for AFM1 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 AFM 1-UPT-LF for milk powder and milk makes possible on-site rapid detection of AFM1 in dairy products quantitatively .