Development of plasmonic biosensors combining reliability and ease of use is still a challenge. Gold nanoparticle arrays made by block copolymer micelle nanolithography (BCMN) stand out for their scalability, cost-effectiveness and tunable plasmonic properties, making them ideal substrates for fluorescence enhancement. Here, we describe a plasmon-enhanced fluorescence immunosensor for the specific and ultrasensitive detection of Plasmodium falciparum lactate dehydrogenase (PfLDH)—a malaria marker—in whole blood. Analyte recognition is realized by oriented antibodies immobilized in a close-packed configuration via the photochemical immobilization technique (PIT), with a top bioreceptor of nucleic acid aptamers recognizing a different surface of PfLDH in a sandwich conformation. The combination of BCMN and PIT enabled maximum control over the nanoparticle size and lattice constant as well as the distance of the fluorophore from the sensing surface. The device achieved a limit of detection smaller than 1 pg/mL (<30 fM) with very high specificity without any sample pretreatment. This limit of detection is several orders of magnitude lower than that found in malaria rapid diagnostic tests or even commercial ELISA kits. Thanks to its overall dimensions, ease of use and high-throughput analysis, the device can be used as a substrate in automated multi-well plate readers and improve the efficiency of conventional fluorescence immunoassays.

结合可靠性和易用性的等离子生物传感器的开发仍然是一个挑战。通过嵌段共聚物胶束纳米光刻技术（BCMN）制成的金纳米颗粒阵列以其可扩展性，成本效益和可调谐的等离激元性质而著称，使其成为荧光增强的理想基材。在这里，我们描述了一种用于全血中恶性疟原虫乳酸脱氢酶（Pf LDH）（一种疟疾标志物）的特异和超灵敏检测的等离激元增强型荧光免疫传感器。通过光化学固定技术（PIT）以紧密包装的形式固定的定向抗体实现了分析物识别，其中核酸适体的顶部生物受体可以识别Pf的不同表面LDH呈三明治构型。BCMN和PIT的组合可以最大程度地控制纳米粒子的大小和晶格常数，以及荧光团与传感表面的距离。该设备无需进行任何样品预处理即可实现非常高的特异性，检测限小于1 pg / mL（<30 fM）。该检测极限比疟疾快速诊断测试甚至商用ELISA试剂盒的检测极限低几个数量级。由于其整体尺寸，易用性和高通量分析，该设备可用作自动化多孔板读取器中的底物，并提高了常规荧光免疫测定的效率。