One-Step Nucleic Acid Purification and Noise-Resistant Polymerase Chain Reaction by Electrokinetic Concentration for Ultralow-Abundance Nucleic Acid Detection.
disease diagnosis
electrokinetics
microfluidics
nucleic acids
polymerase chain reaction
Journal
Angewandte Chemie (International ed. in English)
ISSN: 1521-3773
Titre abrégé: Angew Chem Int Ed Engl
Pays: Germany
ID NLM: 0370543
Informations de publication
Date de publication:
26 06 2020
26 06 2020
Historique:
received:
10
12
2019
revised:
01
04
2020
pubmed:
5
4
2020
medline:
18
3
2021
entrez:
5
4
2020
Statut:
ppublish
Résumé
Nucleic acid amplification tests (NAATs)integrated on a chip hold great promise for point-of-care diagnostics. Currently, nucleic acid (NA) purification remains time-consuming and labor-intensive, and it takes extensive efforts to optimize the amplification chemistry. Using selective electrokinetic concentration, we report one-step, liquid-phase NA purification that is simpler and faster than conventional solid-phase extraction. By further re-concentrating NAs and performing polymerase chain reaction (PCR) in a microfluidic chamber, our platform suppresses non-specific amplification caused by non-optimal PCR designs. We achieved the detection of 5 copies of M. tuberculosis genomic DNA (equaling 0.3 cell) in real biofluids using both optimized and non-optimal PCR designs, which is 10- and 1000-fold fewer than those of the standard bench-top method, respectively. By simplifying the workflow and shortening the development cycle of NAATs, our platform may find use in point-of-care diagnosis.
Identifiants
pubmed: 32246546
doi: 10.1002/anie.201915788
pmc: PMC7560970
mid: NIHMS1621652
doi:
Substances chimiques
DNA, Bacterial
0
Nucleic Acids
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
10981-10988Subventions
Organisme : NIH HHS
ID : P30-ES002109
Pays : United States
Organisme : NIH HHS
ID : R01AI117043
Pays : United States
Organisme : NIAID NIH HHS
ID : U19 AI109755
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI117043
Pays : United States
Organisme : NIEHS NIH HHS
ID : P30 ES002109
Pays : United States
Organisme : NIH HHS
ID : U19AI109755
Pays : United States
Informations de copyright
© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
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