A microfluidic platform integrated with two generated monoclonal antibodies (mAbs) was innovatively fabricated for ultrasensitive detection of enterovirus 71 (EV71). To date, this is the first report on EV71 detection using a microfluidic chip-based immunoassay. The mAbs 1F4 and 2H2 against the major capsid protein VP1 of EV71 were conjugated with carboxyl-functionalized magnetic beads (MBs) and carboxyl-functionalized quantum dots (QDs), respectively. MB-conjugated mAbs separated VP1 from samples, while QD-conjugated mAbs provided fluorescence detection signals and formed a sandwich structure. The monolayer of MBs and QDs captured the VP1 sandwich structure showed stronger fluorescence signal in the microfluidic channel, which could be directly evaluated by fluorescence intensity in response to 365 nm ultraviolet (UV) excitation. The limit of detection (LOD) of the microfluidic platform for VP1 was 10 pg mL⁻¹, which was a 31-fold increase compared to sandwich enzyme-linked immunosorbent assay (ELISA, 310 pg mL⁻¹) using identical antibodies. The microfluidic platform performed well with clinical samples, especially for fecal samples with complex compositions and impurities. The assay is quick to perform (within 30 min), and apparently requires just one syringe without other complex liquid flow system, making it is easy to operate and more suitable for on-site detection and future commercial application.

创新地制造了一个集成了两种生成的单克隆抗体 (mAb) 的微流体平台，用于肠道病毒 71 (EV71) 的超灵敏检测。迄今为止，这是使用基于微流控芯片的免疫测定法检测 EV71 的第一份报告。针对 EV71 的主要衣壳蛋白 VP1 的 mAb 1F4 和 2H2 分别与羧基功能化磁珠 (MB) 和羧基功能化量子点 (QD) 结合。MB 偶联的 mAb 将 VP1 从样品中分离出来，而 QD 偶联的 mAb 提供荧光检测信号并形成夹心结构。MBs 和 QDs 的单层捕获 VP1 夹心结构在微流体通道中显示出更强的荧光信号，可以通过响应 365 nm 紫外线 (UV) 激发的荧光强度直接评估。^-1，与使用相同抗体的夹心酶联免疫吸附测定（ELISA，310 pg mL ^-1）相比，增加了 31 倍。微流控平台在处理临床样本时表现良好，尤其是对于含有复杂成分和杂质的粪便样本。该检测快速执行（30 分钟内），显然只需要一个注射器，无需其他复杂的液流系统，使其易于操作，更适合现场检测和未来的商业应用。