CAJ | 학술논문

建立了一种基于表面等离子体共振( SPR)技术的大肠杆菌特异性快速检测方法。采用EDC/NHS将葡聚糖修饰的CM5芯片表面活化,通过亲和素固定生物素标记的二抗(羊抗兔IgG抗体),利用一抗(兔抗大肠杆菌ATCC25922单克隆抗体)和二抗反应,将兔抗大肠杆菌单克隆抗体固定在传感芯片表面。利用一抗和二抗的质量扩增效应和生物素-亲和素的多级放大效应,实现了对低浓度大肠杆菌ATCC25922的快速检测,提高了SPR传感器灵敏度。利用NaOH溶液对芯片再生,可对多个不同浓度样品进行检测,采用相对响应单位( RU)记录数据。本传感芯片对大肠杆菌ATCC25922响应的线性范围为1．5×102 CFU/mL~1.5×107 CFU/mL,检测限为1.5×102 CFU/mL,相关系数r为0.9815。这种方法简便快捷,有望成为一种在线检测治病菌的有力手段。
건립료일충기우표면등리자체공진( SPR)기술적대장간균특이성쾌속검측방법。채용EDC/NHS장포취당수식적CM5심편표면활화,통과친화소고정생물소표기적이항(양항토IgG항체),이용일항(토항대장간균ATCC25922단극륭항체)화이항반응,장토항대장간균단극륭항체고정재전감심편표면。이용일항화이항적질량확증효응화생물소-친화소적다급방대효응,실현료대저농도대장간균ATCC25922적쾌속검측,제고료SPR전감기령민도。이용NaOH용액대심편재생,가대다개불동농도양품진행검측,채용상대향응단위( RU)기록수거。본전감심편대대장간균ATCC25922향응적선성범위위1．5×102 CFU/mL~1.5×107 CFU/mL,검측한위1.5×102 CFU/mL,상관계수r위0.9815。저충방법간편쾌첩,유망성위일충재선검측치병균적유력수단。
A surface plasmon resonance ( SPR ) biosensor has been established for rapid detection of Escherichia coli. An activation step performed on CM5 chips was carried using EDC/NHS. The biotin-labeled secondard antibody( goat anti-rabbit IgG) was immobilised onto CM5 chip modified with streptavidin via the biotin-streptavidin interaction. Then,rabbit anti-E. coli ATCC25922 polyclonal antibody( primary antibody) was immobilized on sensor chip by antibody-antigen recognition. The detectable signal was amplified by the amplification of primary and secondary antibodies and the streptavidin-biotin amplification strategy. All the SPR experiments were performed used a Biacore 3000 and CM5 chips. Regeneration was achieved using NaOH in order to detect several samples. The change of RU was linearly correlated with the concentration of E. coli ATCC25922(r=0. 981 5). This method exhibited high performance with a dynamic range of 1. 5 × 102 ~1. 5 × 107 CFU/mL, and a detection limit was 1. 5×102 CFU/mL. This method has exhibited great potential in bacteriological testing.