A disposable,integrated loop-mediated isothermal amplification cassette with thermally actuated valves |
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Authors: | Changchun?Liu Michael?G?Mauk Email author" target="_blank">Haim?H?BauEmail author |
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Affiliation: | (1) Department of Mechanical Engineering and Applied Mechanics, University of Pennsylvania, 229 Towne Building, 220 S. 33rd St., Philadelphia, PA 19104-6315, USA; |
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Abstract: | An inexpensive, disposable, integrated, polymer-based cassette for loop-mediated isothermal amplification (LAMP) of target
nucleic acids was designed, fabricated, and tested. The LAMP chamber was equipped with single-use, thermally actuated valves
made with a composite consisting of a mixture of PDMS and expandable microspheres. The effect of the composite composition
on its expansion was investigated, and the valve’s performance was evaluated. In its closed state, the valve can hold pressures
as high as 200 kPa without any significant leakage. Both the LAMP chamber and the valves were actuated with thin film heaters.
The utility of the cassette was demonstrated by carrying out LAMP of Escherichia coli DNA target and reverse transcribed loop meditated isothermal amplification (RT-LAMP) of RNA targets. The amplicons were detected
in real time with a portable, compact detector. The system was capable of detecting as few as 10 target molecules per sample
in well under 1 h. The portable, integrated cassette system described here is particularly suited for applications at the
point of care and in resource-poor countries, where funds and trained personnel are in short supply. |
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