Journal
ANALYTICA CHIMICA ACTA
Volume 1055, Issue -, Pages 65-73Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.aca.2018.12.029
Keywords
Digital polymerase chain reaction; Self-digitization; Vacuum accumulator; Hydration reservoir; Microfluidics
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Funding
- National Natural Science Foundation of China [61771078, 21827812, 81400541]
- Chongqing Research Program of Basic Research and Frontier Technology [cstc2017jcyjBX0036, cstc2016jcyjA0103]
- Chongqing Technical Innovation and Application Demonstration Program [cstc2018jscx-mszdX0073]
- Fundamental Research Funds for the Central Universities [106112016CDJXZ238826]
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We present a self-discretization and zero-water-loss microfluidic digital PCR (dPCR) device to enable low-cost and robust quantitative nucleic acid assays. In this device, a thin void is integrated beneath the reaction chamber array. By utilizing the permeability of polydimethylsiloxane (PDMS) film, the integrated void serves a dual function: vacuum accumulator and hydration reservoir. The combination of pre-stored pumping energy and water compensation for evaporation loss enables simple, robust and reliable single-DNA-molecule amplification and detection in 10,000 reactors of picoliter volume. Compared to the conventional degassing PDMS pumps, the vacuum accumulator exhibits superior performance due to more vacuum storage and shorter diffusion distance. We also evaluated the performance of the embedded hydration layer in suppressing evaporation loss at elevated temperatures, and verified that zero-water-loss could be achieved for all reaction chambers in our dPCR chip during thermal cycling. By performing quantitative detection of T790M DNA from 10 to 10(4) copies/mL, the proposed dPCR chip demonstrated high accuracy and excellent performance for the absolute quantification of the target gene with a dynamic range of 10(4). The simplicity and robustness of our dPCR chip make it well suited for low-cost molecular diagnostic assays under resource-limited settings. (C) 2018 Elsevier B.V. All rights reserved.
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