Highly multiplexed rapid DNA detection with single-nucleotide specificity via convective PCR in a portable device.
Nat Biomed Eng
; 5(7): 702-712, 2021 07.
Article
en En
| MEDLINE
| ID: mdl-34211146
ABSTRACT
Assays for the molecular detection of nucleic acids are typically constrained by the level of multiplexing (this is the case for the quantitative polymerase chain reaction (qPCR) and for isothermal amplification), turnaround times (as with microarrays and next-generation sequencing), quantification accuracy (isothermal amplification, microarrays and nanopore sequencing) or specificity for single-nucleotide differences (microarrays and nanopore sequencing). Here we show that a portable and battery-powered PCR assay performed in a toroidal convection chamber housing a microarray of fluorescently quenched oligonucleotide probes allows for the rapid and sensitive quantification of multiple DNA targets with single-nucleotide discrimination. The assay offers a limit of detection of 10 DNA copies within 30 min of turnaround time and a dynamic range spanning 4 orders of magnitude of DNA concentration, and we show its performance by detecting 20 genomic loci and 30 single-nucleotide polymorphisms in human genomic DNA samples, and 15 bacterial species in clinical isolates. Portable devices for the fast and highly multiplexed detection of nucleic acids may offer advantages in point-of-care diagnostics.
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MEDLINE
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ADN
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Reacción en Cadena de la Polimerasa
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Nat Biomed Eng
Año:
2021
Tipo del documento:
Article