纳米金标记抗体增强SPR检测大肠杆菌O157∶H7  

Immunoanalysis of E.coli O157∶H7 Based on Au Nanoparticles Labelling Antibody Using SPR Biosensor

作　　者：刘霞[1] 李蓉卓[1] 李蕾[1] 李文进 周春娇[2] 

机构地区：[1]湖南农业大学食品科技学院食品科学与生物技术湖南省重点实验室 [2]湖南农业大学理学院,长沙410128

出　　处：《高等学校化学学报》2013年第6期1333-1338,共6页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:31201375);湖南省教育厅优秀青年项目(批准号:10B050)资助

摘　　要：将金纳米粒子(AuNPs)标记的大肠杆菌O157∶H7(E.coli O157∶H7)的多克隆抗体(PAb)作为二抗,采用氨基偶联法将PAb固定在传感器表面作为一抗,通过三明治方法用双通道表面等离子体子共振(SPR)传感器对E.coli O157∶H7进行检测,并与SPR直接法检测进行了比较.结果表明,直接法的检出限为103cfu/mL,线性范围为103～109cfu/mL;AuNPs增强三明治法的检出限为10 cfu/mL,线性范围为10～1010cfu/mL,灵敏度比直接法提高了100倍,且具有更宽的检测范围.本方法不仅检测时间短,而且具有良好的选择性和重现性.The anti-E.coli O157∶ H7 polyclonal antibodies（PAb） were labelled with Au nanoparticles（AuNPs）,which was used as secondary antibodies.The PAb,which was used as primary antibodies,was immobilized on the sensor surface with amino coupling method.The E.coli O157∶ H7 was detected using direct assay and enhancing sandwich assay based on the two channels surface plasmon resonance（SPR） biosensor.The result indicated a good linear relationship between the SPR signal and logarithmic value of E.coli O157∶ H7 concentration ranging from 10^3 cfu/mL to 10^9 cfu/mL,with the limit of detection（LOD） of 10^3 cfu/mL for the direct assay.For AuNPs enhancing sandwich assay,the linear range and the limit of detection were determined to be 10—1010 cfu/mL and 10 cfu/mL,respectively.The sensitivity is 100 times higher than that of direct detection.By introducing AuNPs-PAb compound,a wider detection range and lower detection limit could be obtained.This method is rapid,and has good selectivity and reproducibility.

关 键 词：金纳米粒子 大肠杆菌O157∶H7 表面等离子体子共振 标记 检测 

分 类 号：O657[理学—分析化学]