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From the third issue of 2017, Microarrays has changed its name to High-Throughput.

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Microarrays 2015, 4(4), 474-489; doi:10.3390/microarrays4040474

Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification

School of Engineering and Applied Sciences, University of Pennsylvania, 220 S 33 rd Street, Philadelphia, PA 19104, USA
These authors contributed equally to this work.
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Author to whom correspondence should be addressed.
Academic Editor: Zvi Loewy
Received: 2 August 2015 / Revised: 2 October 2015 / Accepted: 10 October 2015 / Published: 20 October 2015
(This article belongs to the Special Issue Microfluidics Technology)
View Full-Text   |   Download PDF [3869 KB, uploaded 20 October 2015]   |  

Abstract

Microfluidic components and systems for rapid (<60 min), low-cost, convenient, field-deployable sequence-specific nucleic acid-based amplification tests (NAATs) are described. A microfluidic point-of-care (POC) diagnostics test to quantify HIV viral load from blood samples serves as a representative and instructive example to discuss the technical issues and capabilities of “lab on a chip” NAAT devices. A portable, miniaturized POC NAAT with performance comparable to conventional PCR (polymerase-chain reaction)-based tests in clinical laboratories can be realized with a disposable, palm-sized, plastic microfluidic chip in which: (1) nucleic acids (NAs) are extracted from relatively large (~mL) volume sample lysates using an embedded porous silica glass fiber or cellulose binding phase (“membrane”) to capture sample NAs in a flow-through, filtration mode; (2) NAs captured on the membrane are isothermally (~65 °C) amplified; (3) amplicon production is monitored by real-time fluorescence detection, such as with a smartphone CCD camera serving as a low-cost detector; and (4) paraffin-encapsulated, lyophilized reagents for temperature-activated release are pre-stored in the chip. Limits of Detection (LOD) better than 103 virons/sample can be achieved. A modified chip with conduits hosting a diffusion-mode amplification process provides a simple visual indicator to readily quantify sample NA template. In addition, a companion microfluidic device for extracting plasma from whole blood without a centrifuge, generating cell-free plasma for chip-based molecular diagnostics, is described. Extensions to a myriad of related applications including, for example, food testing, cancer screening, and insect genotyping are briefly surveyed. View Full-Text
Keywords: microfluidics; lab on a chip; isothermal nucleic acid amplification microfluidics; lab on a chip; isothermal nucleic acid amplification
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Mauk, M.G.; Liu, C.; Song, J.; Bau, H.H. Integrated Microfluidic Nucleic Acid Isolation, Isothermal Amplification, and Amplicon Quantification. Microarrays 2015, 4, 474-489.

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