基于上转发光免疫层析技术快速检测单核细胞增生李斯特菌  

An up-converting phosphor technology based lateral flow assay for rapid detection of Listeria monocytogenes

作　　者：刘旭[1,2] 李春凤[1,3] 王晓英 冯娜[1,2] 杨瑞馥[1] 王成彬[3] 周蕾[1] 

机构地区：[1]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 [2]北京热景生物技术有限公司,北京102600 [3]解放军总医院临床检验科,北京100853 [4]海军总医院内分泌科,北京100048

出　　处：《军事医学》2014年第9期729-732,共4页Military Medical Sciences

基　　金：国家自然科学基金资助项目(81000774);国家863高技术基金资助项目(2013AA032205);国家科技重大专项资助项目(2011ZX10004-001,2012ZX1004801-002-004,2012ZX1004801-004-015)

摘　　要：目的建立可快速检测单核细胞增生李斯特菌（Listeria monocytogenes,LM）的上转发光免疫层析技术（up-converting phosphor technology based lateral flow assay,UPT-LF）,即LM-UPT-LF。方法针对LM特异的p60蛋白制备单克隆抗体,并与上转发光纳米颗粒（up-converting phosphor nanoparticles,UCP-NPs）共价偶联作为结合物,采用双抗体夹心免疫层析模式建立LM-UPT-LF,并对其敏感性与特异性进行评价。结果建立了可快速检测LM的LM-UPT-LF平台,在样本〈10 cfu、10~99 cfu、100~1000 cfu绝对LM污染量条件下可分别在培养20、18、16 h检出阳性。其他13种食源性致病菌在高浓度（109cfu/ml）时无非特异性交叉。结论所建立的LM-UPT-LF操作简便快速,具有良好的敏感性和特异性,适用于一线病原调查与检测。Objective To develop an up-converting phosphor technology-based lateral-flow（ UPT-LF） assay for rapid detection of Listeria monocytogenes,named LM-UPT-LF. Methods Monoclonal antibodies against p60,which was the specific virulence factor of L. monocytogenes,were prepared and covalently conjugated with up-converting phosphor nanoparticles（ UCP-NPs） as bio-label. Then,LM-UPT-LF was established with double-antibody sandwich mode-based LF assay.Detection performance,including sensitivity and specificity,was evaluated. Results The samples with absolute contaminated amount of L. monocytogenes cells 〈10,10- 99,and 100- 1000 cfu could be significantly detected as positive after incubation at 20 h,18 h,and 16 h,respectively. Other 13 kinds of food-borne pathogens with concentration of 109 cfu /ml did not caused any non-specific reaction. Conclusion The established LM-UPT-LF assay could detect L. monocytogenes with high sensitivity,specificity and simplicity and provides an alternative method for food safety control.

关 键 词：上转发光纳米颗粒 上转发光免疫层析技术 单核细胞增生李斯特菌 

分 类 号：R378.1[医药卫生—病原生物学]