上转换免疫层析方法检测牛奶中大肠杆菌O157H7  

The upconversion lateral flow immunoassay for the detection of Escherichia coli O157:H7 in milk

作　　者：黄震[1] 肖小月 熊智娟 黄艳梅 黄鹤[3] 刘成伟[3] 刘道峰[3] HUANG Zhen;XIAO Xiaoyue;XIONG Zhijuan;HUANG Yanmei;HUANG He;LIU Chengwei;LIU Daofeng(State Key Laboratory of Food Science and Technology,Nanchang University,Nanchang 330047,China;Jiangxi YeLi Medical Device Co.,Ltd.,Nanchang 330008,China;Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease,Jiangxi Province Centre for Disease Control and Prevention,Nanchang 330029,China)

机构地区：[1]南昌大学食品科学与技术国家重点实验室,江西南昌330047 [2]江西业力医疗器械有限公司,江西南昌330008 [3]江西省疾病预防控制中心,江西省食源性疾病诊断溯源重点实验室,江西南昌330029

出　　处：《南昌大学学报（理科版）》2019年第6期556-563,共8页Journal of Nanchang University(Natural Science)

基　　金：国家自然科学基金资助项目(31772066);江西省重点研发计划(20192BBGL70053,20192BBG70069);江西省重点实验室计划项目(20171BCD40021);江西省卫健委科技计划(20196001)。

摘　　要：免疫层析方法是最常见的快速检测方法之一,然而灵敏度低和基质耐受能力差限制了其在食源性致病菌检测中的应用。在本研究中,我们制备了上转换纳米材料(Upconversion Nanoparticles,UCNPs),并将其应用于免疫层析平台。由于UCNPs的反斯托克激发特性,所建立的方法能有效解决灵敏度差及机制耐受能力差的问题。通过系统性优化,所建立的免疫层析方法可在4.6×104 CFU·mL^-1~1×107 CFU·mL^-1浓度范围内实现定量检测,检测限为2.8×104 CFU·mL^-1。该方法在实际牛奶样本检测中表现出优良的抗基质干扰能力,其批内回收率为82.7%~105.4%,变异系数为5.8%~13.3%;批间回收率为79.4%~102.5%,变异系数为6.7%~7.5%。方法灵敏度较高,抗基质干扰能力强,不仅能用于大肠杆菌O157:H7的检测,也可被推广应用于其它食源性致病菌检测。Lateral flow immunoassay(LFIA)is one of the most common rapid detection methods.However,the drawbacks of poor sensitivity and matrix tolerance of LFIA limit its application in detection of foodborne pathogen.Here,to overcome these shortcomings,we synthesized a kind of upconversion nanoparticles(UCNPs)and introduced it into the LFIA platform.Due to the anti-Stokes excitation feature of UCNPs,the proposed LFIA can effectively overcome the above defects.With optimal conditions,the proposed LFIA can quantitative detect Escherichia coli O157H7 in the range of 4.6×104 CFU·mL^-1~1×107 CFU·mL^-1 with the limit of detection of 2.8×104 CFU·mL^-1.Moreover,the method exhibits excellent matrix tolerance in milk.The intra-day recovery rate is 82.7%~105.4%with the coefficient of variation of 5.8%~13.3%,and the inter-assay recovery rate is 79.4%~102.5%with the coefficient of variation of 6.7%~7.5%.The method has excellent performance in sensitivity and matrix tolerance,and can be applied not only to the detection of Escherichia coli O157H7,but also to the detection of other foodborne pathogens.

关 键 词：上转换纳米颗粒 免疫层析 牛奶 大肠杆菌O157:H7 

分 类 号：TS207.4[轻工技术与工程—食品科学]