An integrated, self-contained microfluidic cassette for isolation, amplification, and detection of nucleic acids

Dafeng Chen, Michael Mauk, Xianbo Qiu, Changchun Liu, Jitae Kim, Sudhir Ramprasad, Serge Ongagna, William R. Abrams, Daniel Malamud, Paul L A M Corstjens, Haim H. Bau

Research output: Contribution to journalArticle

Abstract

A self-contained, integrated, disposable, sampleto-answer, polycarbonate microfluidic cassette for nucleic acid-based detection of pathogens at the point of care was designed, constructed, and tested. The cassette comprises on-chip sample lysis, nucleic acid isolation, enzymatic amplification (polymerase chain reaction and, when needed, reverse transcription), amplicon labeling, and detection. On-chip pouches and valves facilitate fluid flow control. All the liquids and dry reagents needed for the various reactions are pre-stored in the cassette. The liquid reagents are stored in flexible pouches formed on the chip surface. Dry (RT-)PCR reagents are pre-stored in the thermal cycling, reaction chamber. The process operations include sample introduction; lysis of cells and viruses; solid-phase extraction, concentration, and purification of nucleic acids from the lysate; elution of the nucleic acids into a thermal cycling chamber and mixing with pre-stored (RT-)PCR dry reagents; thermal cycling; and detection. The PCR amplicons are labeled with digoxigenin and biotin and transmitted onto a lateral flow strip, where the target analytes bind to a test line consisting of immobilized avidin-D. The immobilized nucleic acids are labeled with up-converting phosphor (UCP) reporter particles. The operation of the cassette is automatically controlled by an analyzer that provides pouch and valve actuation with electrical motors and heating for the thermal cycling. The functionality of the device is demonstrated by detecting the presence of bacterial B.Cereus, viral armored RNA HIV, and HIV I virus in saliva samples. The cassette and actuator described here can be used to detect other diseases as well as the presence of bacterial and viral pathogens in the water supply and other fluids.

Original languageEnglish (US)
Pages (from-to)705-719
Number of pages15
JournalBiomedical Microdevices
Volume12
Issue number4
DOIs
StatePublished - Aug 2010

Fingerprint

Microfluidics
Nucleic acids
Nucleic Acids
Amplification
Thermal cycling
Hot Temperature
Polymerase Chain Reaction
polycarbonate
Pathogens
Viruses
Point-of-Care Systems
Immobilized Nucleic Acids
Digoxigenin
Avidin
Water Supply
Solid Phase Extraction
Viral RNA
Biotin
Saliva
Heating

Keywords

  • HIV
  • Lab-on-a-chip
  • Microfluidics
  • PCR chip
  • Point-of-care
  • Pouch
  • RT-PCR

ASJC Scopus subject areas

  • Biomedical Engineering
  • Molecular Biology

Cite this

An integrated, self-contained microfluidic cassette for isolation, amplification, and detection of nucleic acids. / Chen, Dafeng; Mauk, Michael; Qiu, Xianbo; Liu, Changchun; Kim, Jitae; Ramprasad, Sudhir; Ongagna, Serge; Abrams, William R.; Malamud, Daniel; Corstjens, Paul L A M; Bau, Haim H.

In: Biomedical Microdevices, Vol. 12, No. 4, 08.2010, p. 705-719.

Research output: Contribution to journalArticle

Chen, D, Mauk, M, Qiu, X, Liu, C, Kim, J, Ramprasad, S, Ongagna, S, Abrams, WR, Malamud, D, Corstjens, PLAM & Bau, HH 2010, 'An integrated, self-contained microfluidic cassette for isolation, amplification, and detection of nucleic acids', Biomedical Microdevices, vol. 12, no. 4, pp. 705-719. https://doi.org/10.1007/s10544-010-9423-4
Chen, Dafeng ; Mauk, Michael ; Qiu, Xianbo ; Liu, Changchun ; Kim, Jitae ; Ramprasad, Sudhir ; Ongagna, Serge ; Abrams, William R. ; Malamud, Daniel ; Corstjens, Paul L A M ; Bau, Haim H. / An integrated, self-contained microfluidic cassette for isolation, amplification, and detection of nucleic acids. In: Biomedical Microdevices. 2010 ; Vol. 12, No. 4. pp. 705-719.
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AU - Ongagna, Serge

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