Portable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification for antibiotic resistance gene ermB in wastewater

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dc.contributor.authorMao, Kang
dc.contributor.authorZhang, Hua
dc.contributor.authorRan, Fang
dc.contributor.authorCao, Haorui
dc.contributor.authorFeng, Rida
dc.contributor.authorDu, Wei
dc.contributor.authorLi, Xiqing
dc.contributor.authorYang, Zhugen
dc.date.accessioned2023-10-25T09:35:37Z
dc.date.available2023-10-25T09:35:37Z
dc.date.issued2023-10-17
dc.description.abstractWastewater is among the main sources of antibiotic resistance genes (ARGs) in the environment, but effective methods to quickly assess ARGs on-site in wastewater are lacking. Here, using the typical ARG ermB as the target, we report a portable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification (LAMP) for the detection of ARGs. Six primers of LAMP and the crRNA of CRISPR/Cas12a were first designed to be preamplification with LAMP and lead Cas12a to recognize the ermB via base pairing. Due to the trans-cleavage activity of CRISPR/Cas12a after amplicon recognition, ssDNA probes modified with reporter molecules were used to implement a visual assay with lateral flow test strips and fluorescence. After a simple nucleic acid extraction with magnetic beads, the constructed biosensor possesses excellent sensitivity and selectivity as low as 2.75 × 103 copies/μL using fluorescence and later flow strips in wastewater. We further evaluated the community-wide prevalence of ermB in wastewater influent and found high mass loads of ermB during different months. This user-friendly and low-cost biosensor is applicable for rapid on-site ARG detection, providing a potential point-of-use method for rapid assessments of ARG abundance in wastewater from large city areas with many wastewater treatment plants and in resource-limited rural areas.en_UK
dc.identifier.citationMao K, Zhang H, Ran F, et al., (2024) Portable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification for antibiotic resistance gene ermB in wastewater, Journal of Hazardous Materials, Volume 462, January 2024, Article Number 132793en_UK
dc.identifier.eissn1873-3336
dc.identifier.issn0304-3894
dc.identifier.urihttps://doi.org/10.1016/j.jhazmat.2023.132793
dc.identifier.urihttps://dspace.lib.cranfield.ac.uk/handle/1826/20442
dc.language.isoenen_UK
dc.publisherElsevieren_UK
dc.rightsAttribution-NonCommercial-NoDerivatives 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectAntibiotic resistance geneen_UK
dc.subjectCRISPR/Cas12aen_UK
dc.subjectLoop-mediated isothermal amplificationen_UK
dc.subjectBiosensoren_UK
dc.subjectWastewateren_UK
dc.titlePortable biosensor combining CRISPR/Cas12a and loop-mediated isothermal amplification for antibiotic resistance gene ermB in wastewateren_UK
dc.typeArticleen_UK

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