Smart Sensors and Integrated Microsystems, Wayne State University, Detroit, Michigan, United States of America.
Department of Biomedical Engineering, Wayne State University College of Engineering, Detroit, Michigan, United States of America.
PLoS One. 2021 Feb 26;16(2):e0247429. doi: 10.1371/journal.pone.0247429. eCollection 2021.
Influenza virus mutates quickly and unpredictably creating emerging pathogenic strains that are difficult to detect, diagnose, and characterize. Conventional tools to study and characterize virus, such as next generation sequencing, genome amplification (RT-PCR), and serological antibody testing, are not adequately suited to rapidly mutating pathogens like Influenza virus where the success of infection heavily depends on the phenotypic expression of surface glycoproteins. Bridging the gap between genome and pathogenic expression remains a challenge. Using sialic acid as a universal Influenza virus binding receptor, a novel virus avidin-biotin complex-based capture coating was developed and characterized that may be used to create future diagnostic and interrogation platforms for viable whole Influenza virus. First, fluorescent FITC probe studies were used to optimize coating component concentrations. Then atomic force microscopy (AFM) was used to profile the surface characteristics of the novel capture coating, acquire topographical imaging of Influenza particles immobilized by the coating, and calculate the capture efficiency of the coating (over 90%) for all four representative human Influenza virus strains tested.
流感病毒快速且不可预测地发生突变,产生难以检测、诊断和表征的新兴致病株。用于研究和表征病毒的传统工具,如下一代测序、基因组扩增(RT-PCR)和血清学抗体检测,并不完全适合像流感病毒这样快速突变的病原体,因为感染的成功在很大程度上取决于表面糖蛋白的表型表达。弥合基因组和致病表达之间的差距仍然是一个挑战。本研究以唾液酸作为流感病毒的通用结合受体,开发并表征了一种新型的基于病毒亲和素-生物素复合物的捕获涂层,该涂层可用于开发未来用于检测活流感全病毒的诊断和检测平台。首先,利用荧光 FITC 探针研究优化了涂层成分的浓度。然后,原子力显微镜(AFM)用于分析新型捕获涂层的表面特性,获取涂层固定的流感病毒粒子的形貌成像,并计算涂层的捕获效率(超过 90%),对所有四种代表性的人流感病毒株进行了测试。