Colorimetric Polymerase Chain Reaction Enabled by a Fast Light-Activated Substrate Chromogenic Detection Platform.

Autor:	Dong T; Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, College of Chemistry, Analytical & Testing Centre, Sichuan University, 29 Wangjiang Road, Chengdu, Sichuan, P. R. China 610064.; Department of Chemistry, Centre for Biotechnology, Brock University, 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada., Mansour H; Department of Chemistry, Centre for Biotechnology, Brock University, 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada.; Department of Cell Biology, National Research Center, Cairo Governorate 12622, Egypt., Hu H; Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, College of Chemistry, Analytical & Testing Centre, Sichuan University, 29 Wangjiang Road, Chengdu, Sichuan, P. R. China 610064., Wang GA; Department of Chemistry, Centre for Biotechnology, Brock University, 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada., Watson CJF; Department of Health Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada., Yousef M; Department of Health Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada., Matamoros G; Department of Health Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada., Sanchez AL; Department of Health Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada., MacNeil AJ; Department of Health Sciences, Brock University, St. Catharines, Ontario L2S 3A1, Canada., Wu P; Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, College of Chemistry, Analytical & Testing Centre, Sichuan University, 29 Wangjiang Road, Chengdu, Sichuan, P. R. China 610064., Li F; Key Laboratory of Green Chemistry & Technology of Ministry of Education, College of Chemistry, College of Chemistry, Analytical & Testing Centre, Sichuan University, 29 Wangjiang Road, Chengdu, Sichuan, P. R. China 610064.; Department of Chemistry, Centre for Biotechnology, Brock University, 1812 Sir Isaac Brock Way, St. Catharines, Ontario L2S 3A1, Canada.
Jazyk:	angličtina
Zdroj:	Analytical chemistry [Anal Chem] 2020 May 05; Vol. 92 (9), pp. 6456-6461. Date of Electronic Publication: 2020 Apr 16.
DOI:	10.1021/acs.analchem.9b05591
Abstrakt:	Miniaturization of nucleic acid tests (NATs) into portable, inexpensive detection platforms may aid disease diagnosis in point-of-care (POC) settings. Colorimetric signals are ideal readouts for portable NATs, and it remains of high demand to develop color readouts that are simple, quantitative, and versatile. Thus motivated, we report a fast light-activated substrate chromogenic polymerase chain reaction (FLASH PCR) that uses DNA intercalating dyes (DIDs) to enable colorimetric nucleic acid detection and quantification. The FLASH system is established on our finding that DID-DNA intercalation can promote the rapid photooxidation of chromogenic substrates through light-induced production of singlet oxygen. Using this principle, we have successfully converted DID-based fluorescent PCR assays into colorimetric FLASH PCR. To demonstrate the practical applicability of FLASH PCR to POC diagnosis, we also fabricated two readout platforms, including a portable electronic FLASH reader and a paper-based FLASH strip. Using the FLASH reader, we were able to detect as low as 60 copies of DNA standards, a limit of detection (LOD) comparable with commercial quantitative PCR. The FLASH strip further enables the reader-free detection of PCR amplicons by converting the colorimetric signal into the visual measurement of distance as a readout. Finally, the practical applicability of the FLASH PCR was demonstrated by the detection and/or quantification of nucleic acid markers in diverse clinical and biological samples.
Databáze:	MEDLINE
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