Paper device combining CRISPR/Cas12a and reverse-transcription loop-mediated isothermal amplification for SARS-CoV-2 detection in wastewater

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dc.contributor.authorCao, Haorui
dc.contributor.authorMao, Kang
dc.contributor.authorRan, Fang
dc.contributor.authorXu, Pengqi
dc.contributor.authorZhao, Yirong
dc.contributor.authorZhang, Xiangyan
dc.contributor.authorZhou, Hourong
dc.contributor.authorYang, Zhugen
dc.contributor.authorZhang, Hua
dc.contributor.authorJiang, Guibin
dc.date.accessioned2022-10-04T15:04:23Z
dc.date.available2022-10-04T15:04:23Z
dc.date.issued2022-08-30
dc.description.abstractWastewater-based surveillance of the COVID-19 pandemic holds great promise; however, a point-of-use detection method for SARS-CoV-2 in wastewater is lacking. Here, a portable paper device based on CRISPR/Cas12a and reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with excellent sensitivity and specificity was developed for SARS-CoV-2 detection in wastewater. Three primer sets of RT-LAMP and guide RNAs (gRNAs) that could lead Cas12a to recognize target genes via base pairing were used to perform the high-fidelity RT-LAMP to detect the N, E, and S genes of SARS-CoV-2. Due to the trans-cleavage activity of CRISPR/Cas12a after high-fidelity amplicon recognition, carboxyfluorescein-ssDNA-Black Hole Quencher-1 and carboxyfluorescein-ssDNA-biotin probes were adopted to realize different visualization pathways via a fluorescence or lateral flow analysis, respectively. The reactions were integrated into a paper device for simultaneously detecting the N, E, and S genes with limits of detection (LODs) of 25, 310, and 10 copies/mL, respectively. The device achieved a semiquantitative analysis from 0 to 310 copies/mL due to the different LODs of the three genes. Blind experiments demonstrated that the device was suitable for wastewater analysis with 97.7% sensitivity and 82% semiquantitative accuracy. This is the first semiquantitative endpoint detection of SARS-CoV-2 in wastewater via different LODs, demonstrating a promising point-of-use method for wastewater-based surveillance.en_UK
dc.identifier.citationCao H, Mao K, Ran F, et al., (2022) Paper device combining CRISPR/Cas12a and reverse-transcription loop-mediated isothermal amplification for SARS-CoV-2 detection in wastewater. Environmental Science and Technology, Volume 56, Issue 18, September 2022, pp. 13245-13253en_UK
dc.identifier.issn0013-936X
dc.identifier.urihttps://doi.org/10.1021/acs.est.2c04727
dc.identifier.urihttps://dspace.lib.cranfield.ac.uk/handle/1826/18514
dc.language.isoenen_UK
dc.publisherAmerican Chemical Societyen_UK
dc.rightsAttribution-NonCommercial 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc/4.0/*
dc.subjectSARS-CoV-2en_UK
dc.subjectCRISPR/Cas12aen_UK
dc.subjectRT-LAMPen_UK
dc.subjectpaper deviceen_UK
dc.subjectwastewateren_UK
dc.titlePaper device combining CRISPR/Cas12a and reverse-transcription loop-mediated isothermal amplification for SARS-CoV-2 detection in wastewateren_UK
dc.typeArticleen_UK

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