A miniature quantitative PCR device for directly monitoring a sample processing on a microfluidic rapid DNA system

Cedric Hurth, Jianing Yang, Matthew Barrett, Carla Brooks, Alan Nordquist, Stanley Smith, Frederic Zenhausern

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

We report a microfluidic device and measurement method to perform real-time PCR (or qPCR) in a miniaturized configuration for on-chip implementation using reaction volumes of less than 20 μL. The qPCR bioreactor is designed as a module to be embedded in an automated sample-in/profile-out system for rapid DNA biometrics or human identification. The PCR mixture is excited with a 505 nm diode-pumped solid-state laser (DPSSL) and the fluorescence build-up is measured using optical fibers directly embedded to the sidewalls of the microfluidic qPCR bioreactor. We discuss manufacturing and operating parameters necessary to adjust the internal surface conditions and temperature profiles of the bioreactor and to optimize the yield and quality of the PCR reaction for the amplification of 62 bp hTERT intron fragments using the commercial Quantifiler® kit (Life Technologies, Carlsbad, CA) commonly accepted for genotyping analysis. We designed a microfluidic device suitable for continuously processing a specimen by efficiently mixing the reagents from the kit to a set volume of DNA template on chip. Our approach relies on a calibration curve for the specific device using control DNA. We successfully applied this method to determine the concentration of genomic DNA extracted from a buccal swab on separate microfluidic devices which are operated upstream the qPCR device and perform buccal swab lysis and buccal DNA extraction. A precise correlation between the amount determined on chip and that obtained using a commercial cycler is demonstrated.

Original languageEnglish (US)
Pages (from-to)905-914
Number of pages10
JournalBiomedical Microdevices
Volume16
Issue number6
DOIs
StatePublished - Oct 30 2014

Fingerprint

Lab-On-A-Chip Devices
Microfluidics
DNA
Cheek
Bioreactors
Equipment and Supplies
Polymerase Chain Reaction
Monitoring
Processing
Biometric Identification
Optical Fibers
Forensic Anthropology
Solid state lasers
Solid-State Lasers
Biometrics
Introns
Calibration
Amplification
Optical fibers
Real-Time Polymerase Chain Reaction

Keywords

  • Fiberoptics
  • Forensics
  • Microfluidics
  • rapid DNA
  • Real-time PCR

ASJC Scopus subject areas

  • Biomedical Engineering
  • Molecular Biology
  • Medicine(all)

Cite this

A miniature quantitative PCR device for directly monitoring a sample processing on a microfluidic rapid DNA system. / Hurth, Cedric; Yang, Jianing; Barrett, Matthew; Brooks, Carla; Nordquist, Alan; Smith, Stanley; Zenhausern, Frederic.

In: Biomedical Microdevices, Vol. 16, No. 6, 30.10.2014, p. 905-914.

Research output: Contribution to journalArticle

Hurth, Cedric ; Yang, Jianing ; Barrett, Matthew ; Brooks, Carla ; Nordquist, Alan ; Smith, Stanley ; Zenhausern, Frederic. / A miniature quantitative PCR device for directly monitoring a sample processing on a microfluidic rapid DNA system. In: Biomedical Microdevices. 2014 ; Vol. 16, No. 6. pp. 905-914.
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