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Accepted (AM)Open Access (License Unspecified), Open
Journal article
An integrated, cellulose membrane-based PCR chamber
Microsystem technologies : sensors, actuators, systems integration, v 21(4), pp 841-850
01 Apr 2015
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
We report the design, fabrication, and testing of a microfluidic device for molecular diagnostics that sites lysis, solid-phase extraction of nucleic acids, polymerase chain reaction (PCR) amplification, and real-time fluorescence detection in a single chamber. This streamlined design considerably simplifies the fabrication and operation of chip-based nucleic acid assays for detection of pathogens and other disease markers. A single 25-mu L PCR chamber in a plastic chip houses a cellulose-based filter membrane (Whatman FTA(A (R))) for extraction of nucleic acids from the sample. Nucleic acids captured on the filter serve as a template for in situ PCR amplification. The single-use (disposable) microfluidics chip mates with a portable instrument that provides double-sided heating of the PCR chamber for rapid thermal cycling, and incorporates, real time fluorescence detection using a low-cost, miniaturized (ESE GmbH) fluorescence reader. We optimized multiple-pass sample loading of the FTA(A (R)) membrane, vacuum drying of membrane, and membrane wash steps to improve extraction efficiency in a microfluidics format. The chip was tested with samples of Bacillus cereus bacterial culture, and showed a limit of detection of similar to 10(3) target cells. The adaptation of this chip to practical microfluidics applications incorporating integrated pouches for storage and fluid actuation, and pre-loading of dry-stored PCR reagents is discussed, along with potential near-term applications.
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Details
- Title
- An integrated, cellulose membrane-based PCR chamber
- Creators
- Xianbo Qiu - Beijing University of Chemical TechnologyMichael G. Mauk - Univ Penn, Sch Engn & Appl Sci, MEAM Dept, Philadelphia, PA 19104 USA
- Publication Details
- Microsystem technologies : sensors, actuators, systems integration, v 21(4), pp 841-850
- Publisher
- Springer Nature
- Number of pages
- 10
- Grant note
- 81371711 / National Nature Science Foundation of China; National Natural Science Foundation of China (NSFC) U01-DE-017855 / National Institute of Health; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA ZZ1329 / Fundamental Research Funds for the Central Universities of China; Fundamental Research Funds for the Central Universities
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Engineering Technology
- Web of Science ID
- WOS:000351279200015
- Scopus ID
- 2-s2.0-84925291182
- Other Identifier
- 991020623911704721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Engineering, Electrical & Electronic
- Materials Science, Multidisciplinary
- Nanoscience & Nanotechnology
- Physics, Applied