Design and optimization of an ultra-sensitive hairpin DNA aptasensor for Salmonella detection

Lee, Jayeon; Jung, Juyeon; Lee, Chang‐Soo; Ha, Tai Hwan

doi:10.1039/c7ra06000a

Cited by 4 publications

(1 citation statement)

References 20 publications

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“…pH, temperature) compared to their protein counterparts (Kasprowicz et al, 2017;Nakano et al, 2017;Nesbitt et al, 1999). Moreover, they are highly flexible and have been utilized for different applications, including detection of heavy metals (Wu et al, 2010;Yun et al, 2016;Zhang et al, 2011), NA (Lu et al, 2017;Wang et al, 2018), and pathogens (Ali et al, 2017;Kim et al, 2018;Lee et al, 2017), amongst many others. Remarkably, the 10-23 core NAzymes with an inherent RNA-DNA hybrid cleavage activity (Santoro and Joyce, 1997) are among the most widely studied NAzymes.…”

Section: Introductionmentioning

confidence: 99%

DNA-only, microwell-based bioassay for multiplex nucleic acid detection with single base-pair resolution using MNAzymes

Safdar

Ven

Lent

et al. 2020

Biosensors and Bioelectronics

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In disease diagnostics, single-and multiplex nucleic acid (NA) detection, with the potential to discriminate mutated strands, is of paramount importance. Current techniques that rely on target amplification or protein-enzyme based signal amplification are highly relevant, yet still plagued by diverse drawbacks including erroneous target amplification, and the limited stability of protein enzymes. As a solution, we present a multicomponent nucleic acid enzymes (MNAzymes)-based system for singleplex and multiplex detection of NA targets in microwells down to fM concentrations, without the need for any target amplification or protein enzymes, while operating at room temperature and with single base-pair resolution. After successful validation of the MNAzymes in solution, their performance was further verified on beads in bulk and in femtoliter-sized microwells. The latter is not only a highly simplified system compared to previous microwell-based bioassays but, with the detection limit of 180 fM, it is to-date the most sensitive NAzymemediated, bead-based approach, that does not rely on target amplification or any additional signal amplification strategies. Furthermore, we demonstrated, for the first time, multiplexed target detection in microwells, both from buffer and nasopharyngeal swab samples, and presented superior single base-pair resolution of this assay. Because of the design flexibility of MNAzymes and direct demonstration in swab samples, this system holds great promise for multiplexed detection in other clinically relevant matrices without the need of any additional NA or protein components. Moreover, these findings open up the potential for the development of next-generation, protein-free diagnostic tools, including digital assays with single-molecule resolution.

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Section: Introductionmentioning

confidence: 99%

DNA-only, microwell-based bioassay for multiplex nucleic acid detection with single base-pair resolution using MNAzymes

Safdar

Ven

Lent

et al. 2020

Biosensors and Bioelectronics

View full text Add to dashboard Cite

show abstract

Sensitive colorimetric aptasensor based on g-C3N4@Cu2O composites for detection of Salmonella typhimurium in food and water

et al. 2021

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A novel AuNPs colorimetric sensor for sensitively detecting viable Salmonella typhimurium based on dual aptamers

Chen

Yang

et al. 2020

Food Control

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Design and optimization of an ultra-sensitive hairpin DNA aptasensor for Salmonella detection

Abstract: A simple scheme using a hairpin DNA aptasensor was established to detect Salmonella Typhimurium (S. Typhimurium).

Cited by 4 publications

References 20 publications

DNA-only, microwell-based bioassay for multiplex nucleic acid detection with single base-pair resolution using MNAzymes

DNA-only, microwell-based bioassay for multiplex nucleic acid detection with single base-pair resolution using MNAzymes

Sensitive colorimetric aptasensor based on g-C3N4@Cu2O composites for detection of Salmonella typhimurium in food and water

A novel AuNPs colorimetric sensor for sensitively detecting viable Salmonella typhimurium based on dual aptamers

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