A colorimetric and electrochemical immunosensor for point-of-care detection of enterovirus 71
A colorimetric and electrochemical immunosensor for point-of-care detection of enterovirus 71
复制标题
用于即时检测肠道病毒 71 的比色和电化学免疫传感器
DOI：
10.1016/j.bios.2017.07.035
复制
发表时间：
2018-01-15
影响因子：
12.6
通讯作者：
Pang, Dai-Wen
中科院分区：
文献类型：
--
作者：
Hou, Yuan-Hui;Wang, Jia-Jia;Pang, Dai-Wen
人类肠道病毒71型（EV 71）是引起儿童手足口病（HFMD）的主要病原体，其床旁检测对于早期诊断和控制相关流行病是迫切需要的。基于双标记磁性纳米珠扩增技术，建立了一种用于即时检测肠道病毒71型的比色电化学免疫传感器。通过将EV 71单克隆抗体（mAb）和辣根过氧化物酶（HRP）同时固定在磁性纳米珠上，制备了双标记磁性纳米珠（DL-MB）。通过在20 μ L中捕获EV 71病毒体，将样品置于mAb修饰的AuNP涂覆的ITO电极上，随后与DL-MB结合，加入TMB和H2 O2，可以通过肉眼直接读出对应于浓度为1.0 ng mL（-1）的EV 71的比色信号，使得有可能对该病毒进行即时检测。此外，氧化TMB在电极上的还原，无需溶液转移，在同一检测池中即可检测到电化学信号，检测限为0.01ng mL（-1）。经临床样品验证，比色和电化学免疫传感器显示与逆转录聚合酶链反应（RT-PCR）结果完全一致。据我们所知，这是第一次用电化学方法检测EV 71。该检测方法具有灵敏度高、比色检测能力强、操作简便等优点，在病毒相关疾病的即时诊断中具有广阔的应用前景。
Point-of-care detection of human enterovirus 71 (EV71), the major pathogen that causes hand, foot, and mouth disease (HFMD) among children, is urgently needed for early diagnosis and control of related epidemics. A colorimetric and electrochemical immunosensor for point-of-care detection of EV71 has been developed based on dual-labeled magnetic nanobeads amplification. The dual-labeled magnetic nanobeads (DL-MBs) are fabricated by simultaneous immobilization of EV71 monoclonal antibody (mAb) and horseradish peroxidase (HRP) on magnetic nanobeads. By capturing EV71 virions in 20 mu L., sample on mAb modified AuNPs-coated ITO electrode and subsequently binding with DL-MBs, with the addition of TMB and H2O2, colorimetric signals corresponding to EV71 with a concentration of 1.0 ng mL(-1) can be directly read out by naked eyes, making it possible towards point-of care detection of the virus. Furthermore, on the reduction of oxidized TMB on the electrode, electrochemical signal can be detected in the same detection cell without solution transfer, with a detection limit of 0.01 ng mL(-1). Validated with clinical samples, the colorimetric and electrochemical immunosensor shows a complete consistency with reverse transcription-polymerase chain reaction (RT-PCR) results. So far as we know, this is the first report on EV71 detection using electrochemical method. The merits of this assay, including high sensitivity, ability for colorimetric detection and easy to operation, ensure a promising future in point-of-care diagnostics of virus related diseases.