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Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in Environmental Waters

Zhu, Yanzhe and Wu, Xunyi and Gu, Alan Y. and Dobelle, Léopold and Cid, Clément A. and Li, Jing and Hoffmann, Michael R. (2022) Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in Environmental Waters. Environmental Science and Technology, 56 (2). pp. 862-873. ISSN 0013-936X. PMCID PMC8751019. doi:10.1021/acs.est.1c04623.

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Since the COVID-19 pandemic is expected to become endemic, quantification of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in ambient waters is critical for environmental surveillance and for early detection of outbreaks. Herein, we report the development of a membrane-based in-gel loop-mediated isothermal amplification (mgLAMP) system that is designed for the rapid point-of-use quantification of SARS-CoV-2 particles in environmental waters. The mgLAMP system integrates the viral concentration, in-assay viral lysis, and on-membrane hydrogel-based RT-LAMP quantification using enhanced fluorescence detection with a target-specific probe. With a sample-to-result time of less than 1 h, mgLAMP successfully detected SARS-CoV-2 below 0.96 copies/mL in Milli-Q water. In surface water, the lowest detected SARS-CoV-2 concentration was 93 copies/mL for mgLAMP, while the reverse transcription quantitative polymerase chain reaction (RT-qPCR) with optimal pretreatment was inhibited at 930 copies/mL. A 3D-printed portable device is designed to integrate heated incubation and fluorescence illumination for the simultaneous analysis of nine mgLAMP assays. Smartphone-based imaging and machine learning-based image processing are used for the interpretation of results. In this report, we demonstrate that mgLAMP is a promising method for large-scale environmental surveillance of SARS-CoV-2 without the need for specialized equipment, highly trained personnel, and labor-intensive procedures.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Zhu, Yanzhe0000-0002-2260-1830
Wu, Xunyi0000-0001-9710-6896
Gu, Alan Y.0000-0001-8095-3634
Cid, Clément A.0000-0002-7293-035X
Li, Jing0000-0003-0639-9422
Hoffmann, Michael R.0000-0001-6495-1946
Additional Information:© 2021 American Chemical Society. This article is made available via the ACS COVID-19 subset for unrestricted RESEARCH re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. Received: July 10, 2021; Revised: November 17, 2021; Accepted: November 17, 2021; Published: December 30, 2021. The authors thank Caltech Biological Imaging Facility for use of the confocal microscope and Dr. Yuxing Yao for assistance with DLS measurements. This work was supported in whole by the Bill & Melinda Gates Foundation OPP1192379, INV-018569, and INV-030223. Author Contributions: The manuscript was written through contributions of all authors. M.R.H., J.L., and C.A.C. acquired funding for this study. M.R.H. and J.L. conceived the concept of this study. J.L. and Y.Z. designed the study. J.L., Y.Z., X.W., L.D., and A.G. performed the experiments. J.L., Y.Z., and A.G. analyzed the data. Y.Z., J.L., and A.G. wrote the manuscript. M.R.H., L.D., and C.A.C. edited the manuscript. All authors approved the manuscript. The authors declare no competing financial interest. Data and materials availability: The manuscript and the Supporting Information contain all the data needed to evaluate the conclusions in the paper. Correspondence and requests for materials should be addressed to J.L. and M.R.H.
Funding AgencyGrant Number
Bill and Melinda Gates FoundationOPP1192379
Bill and Melinda Gates FoundationINV-018569
Bill and Melinda Gates FoundationINV-030223
Subject Keywords:SARS-CoV-2, RT-LAMP, hydrogel, membrane, environmental quantification of SARS-CoV-2 in milliliters of environmental water samples
Issue or Number:2
PubMed Central ID:PMC8751019
Record Number:CaltechAUTHORS:20220103-231324400
Persistent URL:
Official Citation:Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in Environmental Waters. Yanzhe Zhu, Xunyi Wu, Alan Gu, Leopold Dobelle, Clément A. Cid, Jing Li, and Michael R. Hoffmann. Environmental Science & Technology 2022 56 (2), 862-873; DOI: 10.1021/acs.est.1c04623
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:112649
Deposited By: Tony Diaz
Deposited On:03 Jan 2022 17:20
Last Modified:18 Jan 2022 23:10

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