NDLI: Highly specific, multiplexed isothermal pathogen detection with fluorescent aptamer readout.

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Highly specific, multiplexed isothermal pathogen detection with fluorescent aptamer readout.

Content Provider	Europe PMC
Author	Aufdembrink, Lauren M. Khan, Pavana Gaut, Nathaniel J. Adamala, Katarzyna P. Engelhart, Aaron E.
Copyright Year	2020
Abstract	Isothermal, cell-free, synthetic biology-based approaches to pathogen detection leverage the power of tools available in biological systems, such as highly active polymerases compatible with lyophilization, without the complexity inherent to live-cell systems, of which nucleic acid sequence based amplification (NASBA) is well known. Despite the reduced complexity associated with cell-free systems, side reactions are a common characteristic of these systems. As a result, these systems often exhibit false positives from reactions lacking an amplicon. Here we show that the inclusion of a DNA duplex lacking a promoter and unassociated with the amplicon fully suppresses false positives, enabling a suite of fluorescent aptamers to be used as NASBA tags (Apta-NASBA). Apta-NASBA has a 1 pM detection limit and can provide multiplexed, multicolor fluorescent readout. Furthermore, Apta-NASBA can be performed using a variety of equipment, for example, a fluorescence microplate reader, a qPCR instrument, or an ultra-low-cost Raspberry Pi-based 3D-printed detection platform using a cell phone camera module, compatible with field detection.
Page Count	8
ISSN	13558382
Journal	RNA
Volume Number	26
PubMed Central reference number	PMC7430665
Issue Number	9
PubMed reference number	32482894
e-ISSN	14699001
DOI	10.1261/rna.075192.120
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2020-06-01
Access Restriction	Open
Rights License	This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. © 2020 Aufdembrink et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society
Subject Keyword	NASBA T7 RNA polymerase fluorescent aptamer isothermal amplification pathogen detection
Content Type	Text
Resource Type	Article
Subject	Molecular Biology

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