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Download fileFully Integrated Lab-on-a-Disc for Nucleic Acid Analysis of Food-Borne Pathogens
journal contribution
posted on 2014-04-15, 00:00 authored by Tae-Hyeong Kim, Juhee Park, Chi-Ju Kim, Yoon-Kyoung ChoThis paper describes a micro total
analysis system for molecular
analysis of Salmonella, a major food-borne pathogen.
We developed a centrifugal microfluidic device, which integrated the
three main steps of pathogen detection, DNA extraction, isothermal
recombinase polymerase amplification (RPA), and detection, onto a
single disc. A single laser diode was utilized for wireless control
of valve actuation, cell lysis, and noncontact heating in the isothermal
amplification step, thereby yielding a compact and miniaturized system.
To achieve high detection sensitivity, rare cells in large volumes
of phosphate-buffered saline (PBS) and milk samples were enriched
before loading onto the disc by using antibody-coated magnetic beads.
The entire procedure, from DNA extraction through to detection, was
completed within 30 min in a fully automated fashion. The final detection
was carried out using lateral flow strips by direct visual observation;
detection limit was 10 cfu/mL and 102 cfu/mL in PBS and
milk, respectively. Our device allows rapid molecular diagnostic analysis
and does not require specially trained personnel or expensive equipment.
Thus, we expect that it would have an array of potential applications,
including in the detection of food-borne pathogens, environmental
monitoring, and molecular diagnostics in resource-limited settings.