Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

Abstract Microfluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining...

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Autores principales:	Patrick Hardinge, Divesh K. Baxani, Thomas McCloy, James A. H. Murray, Oliver K. Castell
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/15024b4f29cf432bb09925af9823530b
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spelling	oai:doaj.org-article:15024b4f29cf432bb09925af9823530b2021-12-02T11:57:56ZBioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells10.1038/s41598-020-78996-72045-2322https://doaj.org/article/15024b4f29cf432bb09925af9823530b2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78996-7https://doaj.org/toc/2045-2322Abstract Microfluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining loop-mediated isothermal nucleic acid amplification (LAMP) and bioluminescent assay in real time (BART), with droplet microfluidics, should enable high-throughput, low copy, sequence-specific DNA detection by simple light emission. Stable, uniform LAMP–BART droplets are generated with low cost equipment. The composition and scale of these droplets are controllable and the bioluminescent output during DNA amplification can be imaged and quantified. Furthermore these droplets are readily incorporated into encapsulated droplet interface bilayers (eDIBs), or artificial cells, and the bioluminescence tracked in real time for accurate quantification off chip. Microfluidic LAMP–BART droplets with high stability and uniformity of scale coupled with high throughput and low cost generation are suited to digital DNA quantification at low template concentrations and volumes, where multiple measurement partitions are required. The triggerable reaction in the core of eDIBs can be used to study the interrelationship of the droplets with the environment and also used for more complex chemical processing via a self-contained network of droplets, paving the way for smart soft-matter diagnostics.Patrick HardingeDivesh K. BaxaniThomas McCloyJames A. H. MurrayOliver K. CastellNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Patrick Hardinge Divesh K. Baxani Thomas McCloy James A. H. Murray Oliver K. Castell Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
description	Abstract Microfluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining loop-mediated isothermal nucleic acid amplification (LAMP) and bioluminescent assay in real time (BART), with droplet microfluidics, should enable high-throughput, low copy, sequence-specific DNA detection by simple light emission. Stable, uniform LAMP–BART droplets are generated with low cost equipment. The composition and scale of these droplets are controllable and the bioluminescent output during DNA amplification can be imaged and quantified. Furthermore these droplets are readily incorporated into encapsulated droplet interface bilayers (eDIBs), or artificial cells, and the bioluminescence tracked in real time for accurate quantification off chip. Microfluidic LAMP–BART droplets with high stability and uniformity of scale coupled with high throughput and low cost generation are suited to digital DNA quantification at low template concentrations and volumes, where multiple measurement partitions are required. The triggerable reaction in the core of eDIBs can be used to study the interrelationship of the droplets with the environment and also used for more complex chemical processing via a self-contained network of droplets, paving the way for smart soft-matter diagnostics.
format	article
author	Patrick Hardinge Divesh K. Baxani Thomas McCloy James A. H. Murray Oliver K. Castell
author_facet	Patrick Hardinge Divesh K. Baxani Thomas McCloy James A. H. Murray Oliver K. Castell
author_sort	Patrick Hardinge
title	Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
title_short	Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
title_full	Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
title_fullStr	Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
title_full_unstemmed	Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells
title_sort	bioluminescent detection of isothermal dna amplification in microfluidic generated droplets and artificial cells
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/15024b4f29cf432bb09925af9823530b
work_keys_str_mv	AT patrickhardinge bioluminescentdetectionofisothermaldnaamplificationinmicrofluidicgenerateddropletsandartificialcells AT diveshkbaxani bioluminescentdetectionofisothermaldnaamplificationinmicrofluidicgenerateddropletsandartificialcells AT thomasmccloy bioluminescentdetectionofisothermaldnaamplificationinmicrofluidicgenerateddropletsandartificialcells AT jamesahmurray bioluminescentdetectionofisothermaldnaamplificationinmicrofluidicgenerateddropletsandartificialcells AT oliverkcastell bioluminescentdetectionofisothermaldnaamplificationinmicrofluidicgenerateddropletsandartificialcells
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Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

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