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

Hardinge, Patrick, Baxani, Divesh K., McCloy, Thomas, Murray, James A. H. and Castell, Oliver K. 2020. Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells. Scientific Reports 10 (1) , 21886. 10.1038/s41598-020-78996-7

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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.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Pharmacy
Engineering
Biosciences
Additional Information: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Publisher: Nature Publishing Group
ISSN: 2045-2322
Funders: Wellcome Trust
Date of First Compliant Deposit: 6 January 2021
Date of Acceptance: 2 December 2020
Last Modified: 13 Jan 2021 15:22
URI: http://orca.cf.ac.uk/id/eprint/137373

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