A paper-based colorimetric molecular test for SARS-CoV-2 in saliva

Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging pub...

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Published inBiosensors and bioelectronics. X Vol. 9; p. 100076
Main Authors Davidson, Josiah Levi, Wang, Jiangshan, Maruthamuthu, Murali Kannan, Dextre, Andres, Pascual-Garrigos, Ana, Mohan, Suraj, Putikam, Sai Venkata Sravan, Osman, Fujr Osman Ibrahim, McChesney, Darby, Seville, Jordan, Verma, Mohit S.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 01.12.2021
The Authors. Published by Elsevier B.V
Elsevier
Subjects
Colorimetric LAMP
Microfluidic paper-based analytical devices
Paper-based diagnostics
Saliva
SARS-CoV-2
Colorimetric LAMP
SARS-CoV-2
Paper-based diagnostics
Microfluidic paper-based analytical devices
Saliva
Online AccessGet full text

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Abstract Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets. •Paper-based device utilizes RT-LAMP to detect SARS-CoV-2.•Extraction-free detection of SARS-CoV-2 in whole saliva with minimal processing.•Primer sets target the orf7a, orf7b, and orf1ab regions of SARS-CoV-2.•Configurable reaction zones enable multiplexed detection of pathogens.
AbstractList Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets.
Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets.Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets.
Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal amplification (LAMP) by producing a colorimetric response visible to the human eye. To demonstrate the utility of this device in emerging public health emergencies, we developed and optimized our device to detect SARS-CoV-2 in human saliva without preprocessing. The resulting device was capable of detecting the virus within 60 min and had an analytical sensitivity of 97% and a specificity of 100% with a limit of detection of 200 genomic copies/μL of patient saliva using image analysis. The device consists of a configurable number of reaction zones constructed of Grade 222 chromatography paper separated by 20 mil polystyrene spacers attached to a Melinex® backing via an ARclean® double-sided adhesive. The resulting device is easily configurable to detect multiple targets and has the potential to detect a variety of pathogens simply by changing the LAMP primer sets. •Paper-based device utilizes RT-LAMP to detect SARS-CoV-2.•Extraction-free detection of SARS-CoV-2 in whole saliva with minimal processing.•Primer sets target the orf7a, orf7b, and orf1ab regions of SARS-CoV-2.•Configurable reaction zones enable multiplexed detection of pathogens.
ArticleNumber 100076
Author Mohan, Suraj
Wang, Jiangshan
Osman, Fujr Osman Ibrahim
Dextre, Andres
Putikam, Sai Venkata Sravan
Maruthamuthu, Murali Kannan
Pascual-Garrigos, Ana
McChesney, Darby
Seville, Jordan
Davidson, Josiah Levi
Verma, Mohit S.
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Keywords Colorimetric LAMP
SARS-CoV-2
Paper-based diagnostics
Microfluidic paper-based analytical devices
Saliva
Language English
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2021 The Authors.
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Authors contributed equally to this work.
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Snippet Herein, we describe the development of a paper-based device to detect nucleic acids of pathogens of interest in complex samples using loop-mediated isothermal...
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SubjectTerms Colorimetric LAMP
Microfluidic paper-based analytical devices
Paper-based diagnostics
Saliva
SARS-CoV-2
Title A paper-based colorimetric molecular test for SARS-CoV-2 in saliva
URI https://dx.doi.org/10.1016/j.biosx.2021.100076
https://www.ncbi.nlm.nih.gov/pubmed/34423284
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