A vertical flow paper-microarray assay with isothermal DNA amplification for detection of Neisseria meningitidis.
Paper-based biosensors offer a promising technology to be used at the point of care, enabled by good performance, convenience and low-cost. In this article, we describe a colorimetric vertical-flow DNA microarray (DNA-VFM) that takes advantage of the screening capability of DNA microarrays in a paper format together with isothermal amplification by means of Recombinase Polymerase Amplification (RPA). Different assay parameters such as hybridization buffer, flow rate, printing buffer and capture probe concentration were optimized. A limit of detection (LOD) of 4.4 nM was achieved as determined by tabletop scanning. The DNA-VFM was applied as a proof of concept for detection of Neisseria meningitidis, a primary cause of bacterial meningitis. The LOD was determined to be between 38 and 2.1 × 106 copies/VFMassay, depending on the choice of DNA capture probes. The presented approach provides multiplex capabilities of DNA microarrays in a paper-based format for future point-of-care applications.
['Rivas|Lourdes|L|', 'Reuterswärd|Philippa|P|', 'Rasti|Reza|R|', 'Herrmann|Björn|B|', 'Mårtensson|Andreas|A|', 'Alfvén|Tobias|T|', 'Gantelius|Jesper|J|', 'Andersson-Svahn|Helene|H|']
D004269:DNA, Bacterial D009345:Neisseria meningitidis D021141:Nucleic Acid Amplification Techniques D020411:Oligonucleotide Array Sequence Analysis D010209:Paper D013696:Temperature
Neisseria meningitidis Paper-based biosensor Microarray Point-of-care Gold nanoparticles