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Multiplexed real-time polymerase chain reaction on a digital microfluidic platform.

Publication ,  Journal Article
Hua, Z; Rouse, JL; Eckhardt, AE; Srinivasan, V; Pamula, VK; Schell, WA; Benton, JL; Mitchell, TG; Pollack, MG
Published in: Anal Chem
March 15, 2010

This paper details the development of a digital microfluidic platform for multiplexed real-time polymerase chain reactions (PCR). Liquid samples in discrete droplet format are programmably manipulated upon an electrode array by the use of electrowetting. Rapid PCR thermocycling is performed in a closed-loop flow-through format where for each cycle the reaction droplets are cyclically transported between different temperature zones within an oil-filled cartridge. The cartridge is fabricated using low-cost printed-circuit-board technology and is intended to be a single-use disposable device. The PCR system exhibited remarkable amplification efficiency of 94.7%. To test its potential application in infectious diseases, this novel PCR system reliably detected diagnostic DNA levels of methicillin-resistant Staphylococcus aureus (MRSA), Mycoplasma pneumoniae , and Candida albicans . Amplification of genomic DNA samples was consistently repeatable across multiple PCR loops both within and between cartridges. In addition, simultaneous real-time PCR amplification of both multiple different samples and multiple different targets on a single cartridge was demonstrated. A novel method of PCR speed optimization using variable cycle times has also been proposed and proven feasible. The versatile system includes magnetic bead handling capability, which was applied to the analysis of simulated clinical samples that were prepared from whole blood using a magnetic bead capture protocol. Other salient features of this versatile digital microfluidic PCR system are also discussed, including the configurability and scalability of microfluidic operations, instrument portability, and substrate-level integration with other pre- and post-PCR processes.

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Published In

Anal Chem

DOI

EISSN

1520-6882

Publication Date

March 15, 2010

Volume

82

Issue

6

Start / End Page

2310 / 2316

Location

United States

Related Subject Headings

  • Polymerase Chain Reaction
  • Mycoplasma pneumoniae
  • Microfluidics
  • Methicillin-Resistant Staphylococcus aureus
  • Equipment Design
  • DNA, Fungal
  • DNA, Bacterial
  • Candida albicans
  • Analytical Chemistry
  • 4004 Chemical engineering
 

Citation

APA
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Hua, Z., Rouse, J. L., Eckhardt, A. E., Srinivasan, V., Pamula, V. K., Schell, W. A., … Pollack, M. G. (2010). Multiplexed real-time polymerase chain reaction on a digital microfluidic platform. Anal Chem, 82(6), 2310–2316. https://doi.org/10.1021/ac902510u
Hua, Zhishan, Jeremy L. Rouse, Allen E. Eckhardt, Vijay Srinivasan, Vamsee K. Pamula, Wiley A. Schell, Jonathan L. Benton, Thomas G. Mitchell, and Michael G. Pollack. “Multiplexed real-time polymerase chain reaction on a digital microfluidic platform.Anal Chem 82, no. 6 (March 15, 2010): 2310–16. https://doi.org/10.1021/ac902510u.
Hua Z, Rouse JL, Eckhardt AE, Srinivasan V, Pamula VK, Schell WA, et al. Multiplexed real-time polymerase chain reaction on a digital microfluidic platform. Anal Chem. 2010 Mar 15;82(6):2310–6.
Hua, Zhishan, et al. “Multiplexed real-time polymerase chain reaction on a digital microfluidic platform.Anal Chem, vol. 82, no. 6, Mar. 2010, pp. 2310–16. Pubmed, doi:10.1021/ac902510u.
Hua Z, Rouse JL, Eckhardt AE, Srinivasan V, Pamula VK, Schell WA, Benton JL, Mitchell TG, Pollack MG. Multiplexed real-time polymerase chain reaction on a digital microfluidic platform. Anal Chem. 2010 Mar 15;82(6):2310–2316.
Journal cover image

Published In

Anal Chem

DOI

EISSN

1520-6882

Publication Date

March 15, 2010

Volume

82

Issue

6

Start / End Page

2310 / 2316

Location

United States

Related Subject Headings

  • Polymerase Chain Reaction
  • Mycoplasma pneumoniae
  • Microfluidics
  • Methicillin-Resistant Staphylococcus aureus
  • Equipment Design
  • DNA, Fungal
  • DNA, Bacterial
  • Candida albicans
  • Analytical Chemistry
  • 4004 Chemical engineering