Genetic analysis of H1N1 influenza virus from throat swab samples in a microfluidic system for point-of-care diagnostics

J Am Chem Soc. 2011 Jun 15;133(23):9129-35. doi: 10.1021/ja203981w. Epub 2011 May 24.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA, Single-Stranded / genetics
  • DNA, Single-Stranded / isolation & purification
  • Electrochemistry / instrumentation*
  • Influenza A Virus, H1N1 Subtype / genetics*
  • Influenza A Virus, H1N1 Subtype / isolation & purification*
  • Limit of Detection
  • Magnetics
  • Microfluidic Analytical Techniques*
  • Pharynx / virology*
  • Point-of-Care Systems*
  • Protein Denaturation
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction
  • Ribonucleoproteins / chemistry
  • Specimen Handling*

Substances

  • DNA, Single-Stranded
  • RNA, Viral
  • Ribonucleoproteins