Lai Hsien-Chih, Wang Chih-Hung, Liou Tong-Miin, Lee Gwo-Bin
Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan 30013.
Lab Chip. 2014 Jun 21;14(12):2002-13. doi: 10.1039/c4lc00187g. Epub 2014 May 12.
The influenza A virus is a notorious pathogen that causes high morbidity, high mortality, and even severe global pandemics. Early and rapid diagnosis of the virus is therefore crucial in preventing and controlling any influenza outbreaks. Recently, novel nucleic acid-based affinity reagents called aptamers have emerged as promising candidates for diagnostic assays as they offer several advantages over antibodies, including in vitro selection, chemical synthesis, thermal stability and relatively low costs. Aptamers with high sensitivity and specificity are generated via Systematic Evolution of Ligands by Exponential Enrichment (SELEX), a process that is currently time-consuming, as well as labor- and resource-intensive. In this study, an integrated microfluidic system was developed and was successfully applied to screen a specific aptamer for the influenza A/H1N1 (InfA/H1N1) virus in an automated and highly efficient manner. The selected aptamer was implemented in a magnetic-bead assay, which demonstrated specific and sensitive detection of the InfA/H1N1 virus, even in biological samples such as throat swabs. Consequently, this specific aptamer presents a promising affinity reagent for clinical diagnosis of InfA/H1N1. This is the first demonstration of screening influenza virus-specific aptamers using the microfluidic SELEX technology, which may be expanded for the rapid screening of aptamers against other pathogens for future biomedical applications.
甲型流感病毒是一种臭名昭著的病原体,可导致高发病率、高死亡率,甚至引发严重的全球大流行。因此,对该病毒进行早期快速诊断对于预防和控制任何流感疫情至关重要。最近,一种名为适配体的新型基于核酸的亲和试剂已成为诊断检测的有前景的候选物,因为它们相对于抗体具有多种优势,包括体外筛选、化学合成、热稳定性和相对较低的成本。具有高灵敏度和特异性的适配体是通过指数富集配体系统进化技术(SELEX)产生的,该过程目前既耗时,又耗费人力和资源。在本研究中,开发了一种集成微流控系统,并成功将其应用于以自动化且高效的方式筛选针对甲型流感病毒H1N1亚型(InfA/H1N1)的特异性适配体。所筛选出的适配体被应用于磁珠检测中,该检测即使在咽喉拭子等生物样本中也能特异性且灵敏地检测到InfA/H1N1病毒。因此,这种特异性适配体为InfA/H1N1的临床诊断提供了一种有前景的亲和试剂。这是首次展示使用微流控SELEX技术筛选流感病毒特异性适配体,该技术可能会扩展用于未来生物医学应用中针对其他病原体的适配体的快速筛选。