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Digital Loop-Mediated Isothermal Amplification on a Commercial Membrane

Lin, Xingyu and Huang, Xiao and Urmann, Katharina and Xie, Xing and Hoffmann, Michael R. (2019) Digital Loop-Mediated Isothermal Amplification on a Commercial Membrane. ACS Sensors, 4 (1). pp. 242-249. ISSN 2379-3694. PMCID PMC6350201. https://resolver.caltech.edu/CaltechAUTHORS:20190103-092418331

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Abstract

In this work, we report digital loop-mediated isothermal amplification (LAMP) or reverse-transcription LAMP (RT-LAMP) on a commercial membrane, without the need for complex chip fabrication or use of specialized equipment. Due to the pore size distribution, the theoretical error for digital LAMP on these membranes was analyzed, using a combination of Random Distribution Model and Multi-volume Theory. A facile peel-off process was developed for effective droplets formation on the commercial track-etched polycarbonate (PCTE) membrane. Each pore functions as an individual nanoreactor for single DNA amplification. Absolute quantification of bacteria genomic DNA was realized with a dynamic range from 11 to 1.1  105 copies/µL. One-step digital RT-LAMP was also successfully performed on the membrane for the quantification of MS2 virus in wastewater. With the introduction of new probes, the positive pores can be easily distinguished from negative ones with 100 times difference in fluorescence intensities. Finally, the cost of a disposable membrane is less than $0.1/piece, which, to the best of our knowledge, is the most inexpensive way to perform digital LAMP. The membrane system offers opportunities for point-of-care users or common laboratories to perform digital quantification, single cell analysis, or other bioassays in an inexpensive, flexible and simplified way.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1021/acssensors.8b01419DOIArticle
https://pubs.acs.org/doi/suppl/10.1021/acssensors.8b01419PublisherSupporting Information
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6350201PubMed CentralArticle
ORCID:
AuthorORCID
Lin, Xingyu0000-0002-0950-0736
Huang, Xiao0000-0002-3737-6939
Xie, Xing0000-0002-2253-0964
Hoffmann, Michael R.0000-0002-0432-6564
Alternate Title:Digital LAMP on a Commercial Membrane
Additional Information:© 2019 American Chemical Society. ACS AuthorChoice - This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. Received: November 15, 2018; Accepted: January 3, 2019; Published: January 3, 2019. The authors acknowledge the financial support provided by the Bill and Melinda Gates Foundation (Grant No. OPP1111252). Author Contributions: All authors have given approval to the final version of the manuscript. The authors declare no competing financial interest.
Funders:
Funding AgencyGrant Number
Bill and Melinda Gates FoundationOPP1111252
Subject Keywords:digital LAMP, membrane, microfluidic, droplets, nucleic acid, paper-based analytical device, PCR
Issue or Number:1
PubMed Central ID:PMC6350201
Record Number:CaltechAUTHORS:20190103-092418331
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190103-092418331
Official Citation:Digital Loop-Mediated Isothermal Amplification on a Commercial Membrane. Xingyu Lin, Xiao Huang, Katharina Urmann, Xing Xie, and Michael R. Hoffmann. ACS Sensors 2019 4 (1), 242-249. DOI: 10.1021/acssensors.8b01419
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:92029
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:03 Jan 2019 17:43
Last Modified:03 Oct 2019 20:40

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