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In Situ Nucleic Acid Amplification and Ultrasensitive Colorimetric Readout in a Paper‐Based Analytical Device Using Silver Nanoplates
Author(s) -
SueaNgam Akkapol,
Choopara Ilada,
Li Shangkun,
Schmelcher Mathias,
Somboonraporn,
Howes Philip D.,
deMello Andrew J.
Publication year - 2021
Publication title -
advanced healthcare materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.288
H-Index - 90
eISSN - 2192-2659
pISSN - 2192-2640
DOI - 10.1002/adhm.202001755
Subject(s) - loop mediated isothermal amplification , primer (cosmetics) , in situ , detection limit , materials science , naked eye , nucleic acid amplification tests , linear range , nanotechnology , chemistry , biology , chromatography , dna , virology , biochemistry , organic chemistry , chlamydia trachomatis
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 10 4 copies ( R 2 = 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.