1. In Situ Nucleic Acid Amplification and Ultrasensitive Colorimetric Readout in a Paper-Based Analytical Device Using Silver Nanoplates
- Author
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Naraporn Somboonna, Andrew J. deMello, Philip D. Howes, Shangkun Li, Ilada Choopara, Mathias Schmelcher, and Akkapol Suea-Ngam
- Subjects
In situ ,Methicillin-Resistant Staphylococcus aureus ,Materials science ,Silver ,Concatemer ,Biomedical Engineering ,Loop-mediated isothermal amplification ,Pharmaceutical Science ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Biomaterials ,chemistry.chemical_compound ,Nucleic Acids ,Color contrast ,Detection limit ,business.industry ,Paper based ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,chemistry ,Molecular Diagnostic Techniques ,Nucleic acid ,Optoelectronics ,Colorimetry ,Naked eye ,0210 nano-technology ,business ,Nucleic Acid Amplification Techniques - Abstract
A rapid, highly sensitive, and quantitative colorimetric paper-based analytical device (PAD) based on silver nanoplates (AgNPls) and loop-mediated isothermal amplification (LAMP) is presented. It is shown that cauliflower-like concatemer LAMP products can mediate crystal etching of AgNPls, with a threefold signal enhancement versus linear dsDNA. Methicillin-resistant Staphylococcus aureus (MRSA), an antimicrobial resistant bacterium that poses a formidable risk with persistently high mortality, is used as a model pathogen. Due to the excellent color contrast provided by AgNPls, the PAD allows qualitative analysis by the naked eye and quantitative analysis using a smartphone camera, with detection limits down to a single copy in just 30 min, and a linear response from 1 to 104 copies (R2 = 0.994). The entire assay runs in situ on the paper surface, which drastically simplifies operation of the device. This is the first demonstration of single copy detection using a colorimetric readout, and the developed PAD shows great promise for translation into an ultrasensitive gene-based point-of-care test for any infectious disease target, via modification of the LAMP primer set.
- Published
- 2020