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A potentiometric immunosensor for enterovirus 71 based on bis-MPA-COOH dendrimer-doped AgCl nanospheres with a silver ion-selective electrode
Analyst ( IF 4.2 ) Pub Date : 2017-11-29 00:00:00 , DOI: 10.1039/c7an01305a
Ai-Li Sun 1, 2, 3, 4
Affiliation  

Herein a new potentiometric immunoassay for the point-of-care detection of enterovirus 71 (EV71) was developed by using a silver (Ag+) ion-selective electrode (ISE). Initially, the carboxylated dendrimer-doped AgCl nanospheres were synthesized by the reverse micelle method. Then the synthesized nanospheres were used to label a polyclonal mouse anti-EV71 antibody via a typical carbodiimide coupling method. The immunoreaction was executed on a monoclonal anti-EV71 antibody-coated microplate by using biofunctional AgCl nanospheres as the detection antibody. With a sandwich-type immunoassay format, the carried AgCl nanospheres could be dissolved in the presence of NH3·H2O, and the released silver ions were determined with an external silver ion-selective electrode. Under optimal conditions, the shift in the potential increased with the increase in the EV71 concentration, in a wide linear range of 0.3–300 ng mL−1, with a detection limit of 0.058 ng mL−1. Intra- and inter-assay relative standard deviations with identical batches were less than 4.15% and 6.15%, respectively. By validating the spiked serum samples, our system shows consistency with the enzyme-linked immunosorbent assay (ELISA) kit.

中文翻译:

基于双-MPA-COOH树枝状大分子掺杂的AgCl纳米球和银离子选择电极的肠道病毒71型电位免疫传感器

在本文中,通过使用银(Ag +)离子选择电极(ISE)开发了一种用于肠道病毒71(EV71)即时检测的新型电位免疫测定法。最初,通过逆胶束法合成了羧基化的树枝状大分子掺杂的AgCl纳米球。然后,通过典型的碳二亚胺偶联方法,将合成的纳米球用于标记多克隆小鼠抗EV71抗体。通过使用生物功能性AgCl纳米球作为检测抗体,在单克隆抗EV71抗体包被的微板上进行免疫反应。采用夹心型免疫分析方法,可以在NH 3 ·H 2存在的情况下溶解携带的AgCl纳米球,然后用外部银离子选择电极测定释放的银离子。在最佳条件下,电位变化随EV71浓度的增加而增加,在0.3–300 ng mL -1的宽线性范围内,检出限为0.058 ng mL -1。同一批次的批内和批间相对标准偏差分别小于4.15%和6.15%。通过验证加标的血清样品,我们的系统显示出与酶联免疫吸附测定(ELISA)试剂盒的一致性。
更新日期:2017-11-29
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