Journal article
An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases
Analyst (London), v 136(10), pp 2069-2076
21 May 2011
PMID: 21455542
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
A simple, point of care, inexpensive, disposable cassette for the detection of nucleic acids extracted from pathogens was designed, constructed, and tested. The cassette utilizes a single reaction chamber for isothermal amplification of nucleic acids. The chamber is equipped with an integrated, flow-through, Flinders Technology Associates (Whatman FTA®) membrane for the isolation, concentration, and purification of DNA and/or RNA. The nucleic acids captured by the membrane are used directly as templates for amplification without elution, thus simplifying the cassette’s flow control. The FTA membrane also serves another critical role—enabling the removal of inhibitors that dramatically reduce detection sensitivity. Thermal control is provided with a thin film heater external to the cassette. The amplification process was monitored in real time with a portable, compact fluorescent reader. The utility of the integrated, single-chamber cassette was demonstrated by detecting the presence of HIV-1 in oral fluids. The HIV RNA was reverse transcribed and subjected to loop-mediated, isothermal amplification (LAMP). A detection limit of less than 10 HIV particles was demonstrated. The cassette is particularly suitable for resource poor regions, where funds and trained personnel are in short supply. The cassette can be readily modified to detect nucleic acids associated with other pathogens borne in saliva, urine, and other body fluids as well as in water and food.
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Details
- Title
- An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases
- Creators
- Changchun Liu - Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 South 33rd St., Philadelphia, PA, 19104-6315, USAEran Geva - Department of Basic Sciences, New York University College of Dentistry, New York, 10010, USAMichael Mauk - Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 South 33rd St., Philadelphia, PA, 19104-6315, USAXianbo Qiu - Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 South 33rd St., Philadelphia, PA, 19104-6315, USAWilliam R Abrams - Department of Basic Sciences, New York University College of Dentistry, New York, 10010, USADaniel Malamud - Department of Basic Sciences, New York University College of Dentistry, New York, 10010, USAKelly Curtis - Centers for Disease Control and Prevention, Atlanta, Georgia, USAS. Michele Owen - Centers for Disease Control and Prevention, Atlanta, Georgia, USAHaim H Bau - Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 South 33rd St., Philadelphia, PA, 19104-6315, USA
- Publication Details
- Analyst (London), v 136(10), pp 2069-2076
- Publisher
- Royal Society of Chemistry
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Engineering Technology
- Web of Science ID
- WOS:000289896000009
- Scopus ID
- 2-s2.0-79955141396
- Other Identifier
- 991014878028604721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Chemistry, Analytical