Department of Cogno-Mechatronics Engineering , Pusan National University , Busan 46241 , Republic of Korea.
BioNano Laboratory, School of Engineering , University of Guelph , Gulph , Ontario N1G 2W1 , Canada.
ACS Appl Mater Interfaces. 2018 Apr 18;10(15):12534-12543. doi: 10.1021/acsami.8b02735. Epub 2018 Apr 9.
Rapid and sensitive detection of influenza virus is of soaring importance to prevent further spread of infections and adequate clinical treatment. Herein, an ultrasensitive colorimetric assay called magnetic nano(e)zyme-linked immunosorbent assay (MagLISA) is suggested, in which silica-shelled magnetic nanobeads (MagNBs) and gold nanoparticles are combined to monitor influenza A virus up to femtogram per milliliter concentration. Two essential strategies for ultrasensitive sensing are designed, i.e., facile target separation by MagNBs and signal amplification by the enzymelike activity of gold nanozymes (AuNZs). The enzymelike activity was experimentally and computationally evaluated, where the catalyticity of AuNZ was tremendously stronger than that of normal biological enzymes. In the spiked test, a straightforward linearity was presented in the range of 5.0 × 10-5.0 × 10g·mL in detecting the influenza virus A (New Caledonia/20/1999) (H1N1). The detection limit is up to 5.0 × 10 g·mL only by human eyes, as well as up to 44.2 × 10 g·mL by a microplate reader, which is the lowest record to monitor influenza virus using enzyme-linked immunosorbent assay-based technology as far as we know. Clinically isolated human serum samples were successfully observed at the detection limit of 2.6 PFU·mL. This novel MagLISA demonstrates, therefore, a robust sensing platform possessing the advances of fathomable sample separation, enrichment, ultrasensitive readout, and anti-interference ability may reduce the spread of influenza virus and provide immediate clinical treatment.
快速灵敏地检测流感病毒对于防止感染进一步传播和进行适当的临床治疗至关重要。在此，提出了一种超灵敏比色分析方法，称为磁性纳米（e）酶联免疫吸附测定（MagLISA），其中将二氧化硅壳层磁性纳米珠（MagNBs）和金纳米颗粒结合在一起，可监测每毫升低至飞克浓度的流感 A 病毒。设计了两种用于超灵敏传感的基本策略，即通过 MagNBs 进行简便的目标分离和通过金纳米酶（AuNZs）的酶样活性进行信号放大。实验和计算评估了酶样活性，其中 AuNZ 的催化活性远远强于正常生物酶。在加标测试中，在检测流感病毒 A（新喀里多尼亚/20/1999）（H1N1）的 5.0×10-5.0×10g·mL 范围内呈现出直接线性关系。仅用人眼就可以达到 5.0×10 g·mL 的检测限，而通过微孔板读数器可以达到 44.2×10 g·mL，这是迄今为止使用基于酶联免疫吸附测定技术监测流感病毒的最低记录。在检测限为 2.6 PFU·mL 时，成功观察到临床分离的人血清样本。因此，这种新型 MagLISA 展示了一种强大的传感平台，具有可理解的样品分离、富集、超灵敏读出和抗干扰能力的优势，可以减少流感病毒的传播并提供即时的临床治疗。
