Journal
THERANOSTICS
Volume 7, Issue 8, Pages 2220-2230Publisher
IVYSPRING INT PUBL
DOI: 10.7150/thno.18675
Keywords
loop-mediated isothermal amplification; paper; biosensor; molecular diagnosis; nucleic acid testing; point-of-care
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Funding
- National Research Foundation of Korea (NRF) [NRF-2013M3C1A8 A01072759]
- R&D Joint Venture Program of the NRF [2015K000184]
- Original Technology Research Program for Brain Science through the National Research Foundation of Korea (NRF) [NRF-2015M3C7A1029196]
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Paper-based diagnostic devices have many advantages as a one of the multiple diagnostic test platforms for point-of-care (POC) testing because they have simplicity, portability, and cost-effectiveness. However, despite high sensitivity and specificity of nucleic acid testing (NAT), the development of NAT based on a paper platform has not progressed as much as the others because various specific conditions for nucleic acid amplification reactions such as pH, buffer components, and temperature, inhibitions from technical differences of paper-based device. Here, we propose a paper-based device for performing loop-mediated isothermal amplification (LAMP) with real-time simultaneous detection of multiple DNA targets. We determined the optimal chemical components to enable dry conditions for the LAMP reaction without lyophilization or other techniques. We also devised the simple paper device structure by sequentially stacking functional layers, and employed a newly discovered property of hydroxynaphthol blue fluorescence to analyze real-time LAMP signals in the paper device. This proposed platform allowed analysis of three different meningitis DNA samples in a single device with single-step operation. This LAMP-based multiple diagnostic device has potential for real-time analysis with quantitative detection of 10(2)-10(5) copies of genomic DNA. Furthermore, we propose the transformation of DNA amplification devices to a simple and affordable paper system approach with great potential for realizing a paper-based NAT system for POC testing.
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