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Article: Integration of RT-LAMP and Microfluidic Technology for Detection of SARS-CoV-2 in Wastewater as an Advanced Point-of-Care Platform

TitleIntegration of RT-LAMP and Microfluidic Technology for Detection of SARS-CoV-2 in Wastewater as an Advanced Point-of-Care Platform
Authors
KeywordsMicrofluidic device
N gene
RT-LAMP
SARS-CoV-2
Issue Date4-May-2022
PublisherSpringer
Citation
Food and Environmental Virology, 2022, v. 14, n. 4, p. 364-373 How to Cite?
Abstract

Abstract: Development of lab-on-a-chip (LOC) system based on integration of reverse transcription loop-mediated isothermal amplification (RT-LAMP) and microfluidic technology is expected to speed up SARS-CoV-2 diagnostics allowing early intervention. In the current work, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and RT-LAMP assays were performed on extracted RNA of seven wastewater samples from COVID-19 hotspots. RT‑LAMP assay was also performed on wastewater samples without RNA extraction. Current detection of SARS-CoV-2 is mainly by RT-qPCR of ORF (ORF1ab) and N genes so we targeted both to find the best target gene for SARS-CoV-2 detection. We also performed RT-LAMP with/without RNA extraction inside microfluidic device to target both genes. Positivity rates of RT-qPCR and RT-LAMP performed on extracted RNA were 100.0% (7/7) and 85.7% (6/7), respectively. RT-qPCR results revealed that all 7 wastewater samples were positive for N gene (Ct range 37–39), and negative for ORF1ab, suggesting that N gene could be the best target gene for SARS-CoV-2 detection. RT-LAMP of N and ORF (ORF1a) genes performed on wastewater samples without RNA extraction indicated that all 7 samples remains pink (negative). The color remains pink in all microchannels except microchannels which subjected to RT-LAMP for targeting N region after RNA extraction (yellow color) in 6 out of 7 samples. This study shows that SARS-CoV-2 was successfully detected from wastewater samples using RT-LAMP in microfluidic chips. This study brings the novelty involving the use of wastewater samples for detection of SARS-CoV-2 without previous virus concentration and with/without RNA extraction.


Persistent Identifierhttp://hdl.handle.net/10722/338885
ISSN
2023 Impact Factor: 4.1
2023 SCImago Journal Rankings: 0.568
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorDonia, A-
dc.contributor.authorFurqan, Shahid M-
dc.contributor.authorHassan, Su-
dc.contributor.authorShahid, R-
dc.contributor.authorAhmad, A-
dc.contributor.authorJaved, A-
dc.contributor.authorNawaz, M-
dc.contributor.authorYaqub, T-
dc.contributor.authorBokhari, H -
dc.date.accessioned2024-03-11T10:32:15Z-
dc.date.available2024-03-11T10:32:15Z-
dc.date.issued2022-05-04-
dc.identifier.citationFood and Environmental Virology, 2022, v. 14, n. 4, p. 364-373-
dc.identifier.issn1867-0334-
dc.identifier.urihttp://hdl.handle.net/10722/338885-
dc.description.abstract<p>Abstract: Development of lab-on-a-chip (LOC) system based on integration of reverse transcription loop-mediated isothermal amplification (RT-LAMP) and microfluidic technology is expected to speed up SARS-CoV-2 diagnostics allowing early intervention. In the current work, reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and RT-LAMP assays were performed on extracted RNA of seven wastewater samples from COVID-19 hotspots. RT‑LAMP assay was also performed on wastewater samples without RNA extraction. Current detection of SARS-CoV-2 is mainly by RT-qPCR of ORF (ORF1ab) and N genes so we targeted both to find the best target gene for SARS-CoV-2 detection. We also performed RT-LAMP with/without RNA extraction inside microfluidic device to target both genes. Positivity rates of RT-qPCR and RT-LAMP performed on extracted RNA were 100.0% (7/7) and 85.7% (6/7), respectively. RT-qPCR results revealed that all 7 wastewater samples were positive for N gene (Ct range 37–39), and negative for ORF1ab, suggesting that N gene could be the best target gene for SARS-CoV-2 detection. RT-LAMP of N and ORF (ORF1a) genes performed on wastewater samples without RNA extraction indicated that all 7 samples remains pink (negative). The color remains pink in all microchannels except microchannels which subjected to RT-LAMP for targeting N region after RNA extraction (yellow color) in 6 out of 7 samples. This study shows that SARS-CoV-2 was successfully detected from wastewater samples using RT-LAMP in microfluidic chips. This study brings the novelty involving the use of wastewater samples for detection of SARS-CoV-2 without previous virus concentration and with/without RNA extraction.</p>-
dc.languageeng-
dc.publisherSpringer-
dc.relation.ispartofFood and Environmental Virology-
dc.rightsThis work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.-
dc.subjectMicrofluidic device-
dc.subjectN gene-
dc.subjectRT-LAMP-
dc.subjectSARS-CoV-2-
dc.titleIntegration of RT-LAMP and Microfluidic Technology for Detection of SARS-CoV-2 in Wastewater as an Advanced Point-of-Care Platform-
dc.typeArticle-
dc.identifier.doi10.1007/s12560-022-09522-3-
dc.identifier.scopuseid_2-s2.0-85129640308-
dc.identifier.volume14-
dc.identifier.issue4-
dc.identifier.spage364-
dc.identifier.epage373-
dc.identifier.eissn1867-0342-
dc.identifier.isiWOS:000790632000001-
dc.identifier.issnl1867-0334-

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