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Inter-laboratory calibration of quantitative analyses of antibiotic resistance genes

2020Journal article Restricted access
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dc.contributor.authorRocha, Jaqueline
dc.contributor.authorCacace, Damiano
dc.contributor.authorKampouris, Ioannis
dc.contributor.authorGuilloteau, Hélène
dc.contributor.authorJäger, Thomas
dc.contributor.authorMarano, Roberto B.M.
dc.contributor.authorKaraolia, Popi
dc.contributor.authorManaia, Célia M.
dc.contributor.authorMerlin, Christophe
dc.contributor.authorFatta-Kassinos, Despo
dc.contributor.authorCytryn, Eddie
dc.contributor.authorBerendonk, Thomas U.
dc.contributor.authorSchwartz, Thomas
dc.date.accessioned2021-02-03T11:04:07Z
dc.date.available2021-02-03T11:04:07Z
dc.date.issued2020
dc.description.abstractAntibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are widely distributed in the environment where they represent potential public health threats. Quantitative PCR (qPCR) is a suitable approach to detect and quantify ARGs in environmental samples. However, the comparison of gene quantification data between different laboratories is challenging since the data are predominantly obtained under non-harmonized conditions, using different qPCR protocols. This study aimed at carrying out an inter-laboratory calibration in order to assess the variability inherent to the qPCR procedures for quantification of ARGs. With this aim, samples of treated wastewater collected in three different countries were analysed based on common DNA extract pools and identical protocols as well as distinct equipment, reagents batches, and operators. The genes analysed were the 16S rRNA, vanA, blaTEM, qnrS, sul1, blaCTXM-32 and intI1 and the artificial pNORM1 plasmid containing fragments from the seven targeted genes was used as a reference. The 16S rRNA gene was the most abundant, in all the analysed samples, followed by intI1, sul1, qnrS, and blaTEM, while blaCTXM-32 and vanA were below the limit of quantification in most or all the samples. For the genes 16S rRNA, sul1, intI1, blaTEM and qnrS the inter-laboratory variation was below 28% (3–8%, 6–18%, 8–21%, 10–24%, 15–28%, respectively). While it may be difficult to fully harmonize qPCR protocols due to equipment, reagents and operator variations, the inter-laboratory calibration is an adequate and necessary step to increase the reliability of comparative data on ARGs abundance in different environmental compartments and/or geographic regions.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationRocha, J., Cacace, D., Kampouris, I., Guilloteau, H., Jäger, T., Marano, R. B., ... Schwartz, T. (2020). Inter-laboratory calibration of quantitative analyses of antibiotic resistance genes. Journal of Environmental Chemical Engineering, 8(1), 102214pt_PT
dc.identifier.doi10.1016/j.jece.2018.02.022pt_PT
dc.identifier.eid85042650412
dc.identifier.eissn2213-3437
dc.identifier.urihttp://hdl.handle.net/10400.14/31868
dc.identifier.wos000515128500007
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevierpt_PT
dc.relationNORTE-08-5369-FSE-000007pt_PT
dc.relation821-0000-13pt_PT
dc.relationANtibioticS and mobile resistance elements in WastEwater Reuse applications: risks and innovative solutions
dc.subjectInter-laboratory calibrationpt_PT
dc.subjectQuantitative PCRpt_PT
dc.subjectWastewaterpt_PT
dc.subjectAntibiotic resistance genept_PT
dc.titleInter-laboratory calibration of quantitative analyses of antibiotic resistance genespt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleANtibioticS and mobile resistance elements in WastEwater Reuse applications: risks and innovative solutions
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/WaterJPI%2F0001%2F2013/PT
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/675530/EU
oaire.citation.issue1pt_PT
oaire.citation.titleJournal of Environmental Chemical Engineeringpt_PT
oaire.citation.volume8pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStreamH2020
person.familyNameRocha
person.familyNameCacace
person.familyNameMarano
person.familyNameKaraolia
person.familyNameManaia
person.familyNameMerlin
person.familyNameFatta-Kassinos
person.givenNameJaqueline
person.givenNameDamiano
person.givenNameRoberto
person.givenNamePopi
person.givenNameCélia
person.givenNameChristophe
person.givenNameDespo
person.identifier565525
person.identifier.ciencia-id721C-C3E4-6A38
person.identifier.ciencia-idC31F-553B-6365
person.identifier.orcid0000-0003-1651-9483
person.identifier.orcid0000-0003-1725-1044
person.identifier.orcid0000-0001-7805-6360
person.identifier.orcid0000-0003-0867-8177
person.identifier.orcid0000-0002-3674-1789
person.identifier.orcid0000-0002-9261-5322
person.identifier.orcid0000-0003-1173-0941
person.identifier.ridK-3850-2014
person.identifier.ridG-2331-2016
person.identifier.scopus-author-id57113035100
person.identifier.scopus-author-id55848052100
person.identifier.scopus-author-id6602465318
person.identifier.scopus-author-id6602188649
person.identifier.scopus-author-id6603838891
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.identifierhttp://doi.org/10.13039/501100008530
project.funder.nameFundação para a Ciência e a Tecnologia
project.funder.nameEuropean Commission
rcaap.rightsrestrictedAccesspt_PT
rcaap.typearticlept_PT
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