Genetic Analysis of H1N1 Influenza Virus from Throat Swab Samples in a Microfluidic System for Point-of-Care Diagnostics,Journal of the American Chemical Society

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Genetic Analysis of H1N1 Influenza Virus from Throat Swab Samples in a Microfluidic System for Point-of-Care Diagnostics
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2011-06-15 , DOI: 10.1021/ja203981w
B Scott Ferguson ₁ , Steven F Buchsbaum , Ting-Ting Wu , Kuangwen Hsieh , Yi Xiao , Ren Sun , H Tom Soh

Affiliation

The ability to obtain sequence-specific genetic information about rare target organisms directly from complex biological samples at the point-of-care would transform many areas of biotechnology. Microfluidics technology offers compelling tools for integrating multiple biochemical processes in a single device, but despite significant progress, only limited examples have shown specific, genetic analysis of clinical samples within the context of a fully integrated, portable platform. Herein we present the Magnetic Integrated Microfluidic Electrochemical Detector (MIMED) that integrates sample preparation and electrochemical sensors in a monolithic disposable device to detect RNA-based virus directly from throat swab samples. By combining immunomagnetic target capture, concentration, and purification, reverse-transcriptase polymerase chain reaction (RT-PCR) and single-stranded DNA (ssDNA) generation in the sample preparation chamber, as well as sequence-specific electrochemical DNA detection in the electrochemical cell, we demonstrate the detection of influenza H1N1 in throat swab samples at loads as low as 10 TCID(50), 4 orders of magnitude below the clinical titer for this virus. Given the availability of affinity reagents for a broad range of pathogens, our system offers a general approach for multitarget diagnostics at the point-of-care.

中文翻译：

在用于即时诊断的微流体系统中对咽拭子样本中的 H1N1 流感病毒进行遗传分析

在护理点直接从复杂的生物样本中获取有关稀有目标生物体的序列特异性遗传信息的能力将改变生物技术的许多领域。微流体技术为在单个设备中集成多个生化过程提供了令人信服的工具，但尽管取得了重大进展，但只有有限的例子显示了在完全集成的便携式平台背景下对临床样本进行特定的遗传分析。在此，我们推出了磁集成微流体电化学检测器 (MIMED)，它将样品制备和电化学传感器集成在一个整体一次性设备中，以直接从咽拭子样品中检测基于 RNA 的病毒。通过将免疫磁性目标捕获、浓缩和纯化、逆转录酶聚合酶链式反应 (RT-PCR) 和样品制备室中的单链 DNA (ssDNA) 生成以及电化学池中的序列特异性电化学 DNA 检测相结合，我们证明了在咽拭子样本中检测到 H1N1 流感病毒的载量低至 10 TCID(50)，比该病毒的临床滴度低 4 个数量级。鉴于亲和试剂适用于多种病原体，我们的系统提供了一种在护理点进行多目标诊断的通用方法。

更新日期：2011-06-15

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