基于上转发光免疫层析技术的常见食源性致病菌快速检测方法研究与评价  

Development of up-converting phosphor technology based lateral flow assay for quantitative detection of foodborne pathogens

作　　者：李春凤[1,2] 赵勇[2] 王晓英 张平平[2] 刘晓[2,4] 孙崇云[1,2] 杨瑞馥[2] 王成彬[1] 周蕾[2] 

机构地区：[1]解放军总医院临床检验科,北京100853 [2]军事医学科学院微生物流行病研究所,病原微生物生物安全国家重点实验室,北京100071 [3]海军总医院内分泌科,北京100048 [4]重庆医科大学公共卫生与管理学院,重庆400016

出　　处：《军事医学》2015年第2期128-132,共5页Military Medical Sciences

基　　金：国家863计划资助项目(2013AA032205);国家科技重大专项资助项目(2011ZX10004-001,2012ZX10004801-002-004,2012ZX10004801-004-015);北京市科委2014年度科技创新基地培育与发展工程专项资助项目(Z141107004414026);国家自然科学基金资助项目(81000774);北京市科技新星资助项目

摘　　要：目的研制针对4种常见食源性致病菌：甲型副伤寒沙门菌（Salmonella paratyphi A）、乙型副伤寒沙门菌（Salmonella paratyphi B）、大肠埃希菌O157∶H7（Escherichia coli O157∶H7）、副溶血弧菌（Vibrio parahaemolyticus）的基于上转发光免疫层析技术（UPT-LF）的快速定量检测试纸,并对其检测性能进行评价。方法以上转发光纳米颗粒（UCP-NPs）作为示踪物,基于双抗体夹心检测模型,研制针对上述4种靶标菌的UPT-LF试纸;以4种靶标菌的系列浓度标准菌悬液作为标准样品,评价试纸的敏感性、特异性、线性和精密性,并进行模拟染菌食品的检测,评价模拟样品阳性检出率。结果针对4种靶标菌的UPT-LF试纸均具有良好的线性,相关系数r为0.985-0.996;检测敏感性达10^5-10^6CFU/ml,与其他近缘菌株无交叉反应;对乳制品、饮料、小食品、水产、肉类等细菌污染的食品检出率较高。结论该研究建立的对4种常见食源性致病菌进行快速检测的UPT-LF,简便快速,具有良好的敏感性、特异性和线性定量能力,操作性能可满足食品安全检测的要求。Objective To develop an up-converting phosphor technology based lateral flow（ UPT-LF） assay for rapid detection of Salmonella paratyphi A, S. paratyphi B, Escherichia coli O157 ∶ H7 and Vibrio parahaemolyticus.Methods With up-converting phosphor nano-particles（ UCP-NPs） as the bio-marker,four double-antibody-sandwich mode based UPT-LF strips for detecting the above mentioned four pathogens were prepared respectively and their sensitivity,accuracy,linearity and specificity were evaluated. Furthermore,the feasibility of detecting bacteria in food samples was evaluated by different food samples artificially contaminated with less than 10 CFU target pathogens. Results The sensitivity of UPT-LF assays for four pathogens was 10^5- 10^6 CFU / ml with excellent specificity. The four strips had a good linear response with the linear fitting coefficient of determination（ r） for each target pathogen ranging from 0. 985 to 0. 996. The positive rate of detecting pathogens from samples was acceptable. Conclusion The four developed UPT-LF strips provide a new choice for rapid,specific and sensitive and quantitative detection of S. paratyphi A,S. paratyphi B,E. coli O157∶ H7 and V. parahemolyticus.

关 键 词：上转发光纳米颗粒 侧流术 定量检测 食源性致病菌 甲型副伤寒沙门菌 乙型副伤寒沙门菌 大肠埃希菌O157∶H7 副溶血弧菌 

分 类 号：R155.5[医药卫生—营养与食品卫生学]