Microfluidic Biosensor for the Serotype-Specific Detection of Dengue Virus RNA

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• 作者：Natalya V. Zaytseva ; Richard A. Montagna ; and Antje J. Baeumner
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：December 1, 2005
• 年：2005
• 卷：77
• 期：23
• 页码：7520 - 7527
• 全文大小：360K
• 年卷期：v.77,no.23(December 1, 2005)
• ISSN：1520-6882

文摘

The development of a microfluidic biosensor with fluorescence detection for the rapid, sensitive, and serotype-specific detection of Dengue virus is presented. Thebiosensor chip consists of poly(dimethylsiloxane) (PDMS)substrate with fabricated microchannels and a glasssubstrate used to seal the microchannels. These twosubstrates are packaged within a pressure-closed Plexiglas housing to provide a watertight reversible sealing atthe PDMS-glass interface. The ability to reversibly sealthe device permits easy disassembly and quick interchange of the device parts, which is ideal for developmental purposes. The biosensor employs a magneticbead-based sandwich hybridization system in conjugationwith liposome amplification for the specific detection ofnucleic acids. The concentrations of the various biosensorcomponents were optimized using a synthesized fragmentof Dengue virus RNA. To evaluate the sensitivity of theassay, two detection systems, based on fluorescencemeasurements of intact and lysed liposomes, were analyzed. The entire analysis was complete within 20 min(including incubation time) with RNA detection limits of0.125 nM and 50 pM for intact and lysed liposomedetection systems, respectively. Subsequently, the biosensor was applied to the analysis of actual RNA obtainedfrom Dengue virus serotypes 1-4. The resulting signalswere compared to those obtained using standard electrochemiluminescence detection and shown to correspondperfectly with respect to serotype identification.