An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases

Changchun Liu, Eran Geva, Michael Mauk, Xianbo Qiu, William R. Abrams, Daniel Malamud, Kelly Curtis, S. Michele Owen, Haim H. Bau

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)2069-2076
Number of pages8
JournalAnalyst
Volume136
Issue number10
DOIs
StatePublished - May 21 2011

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Fingerprint Dive into the research topics of 'An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases'. Together they form a unique fingerprint.

  • Cite this

    Liu, C., Geva, E., Mauk, M., Qiu, X., Abrams, W. R., Malamud, D., Curtis, K., Owen, S. M., & Bau, H. H. (2011). An isothermal amplification reactor with an integrated isolation membrane for point-of-care detection of infectious diseases. Analyst, 136(10), 2069-2076. https://doi.org/10.1039/c1an00007a