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dc.contributor.authorCard, RM
dc.contributor.authorWarburton, Philip
dc.contributor.authorMacLaren, N
dc.contributor.authorMullany, P
dc.contributor.authorAllan, E
dc.contributor.authorAnjum, MF
dc.date.accessioned2015-10-14T11:34:11Z
dc.date.available2015-10-14T11:34:11Z
dc.date.issued2014-01-22
dc.identifier.issn1932-6203
dc.identifier.issn1932-6203
dc.identifier.othere86428
dc.identifier.urihttp://hdl.handle.net/10026.1/3628
dc.description.abstract

The aim of this study was to screen for the presence of antimicrobial resistance genes within the saliva and faecal microbiomes of healthy adult human volunteers from five European countries. Two non-culture based approaches were employed to obviate potential bias associated with difficult to culture members of the microbiota. In a gene target-based approach, a microarray was employed to screen for the presence of over 70 clinically important resistance genes in the saliva and faecal microbiomes. A total of 14 different resistance genes were detected encoding resistances to six antibiotic classes (aminoglycosides, β-lactams, macrolides, sulphonamides, tetracyclines and trimethoprim). The most commonly detected genes were erm(B), blaTEM, and sul2. In a functional-based approach, DNA prepared from pooled saliva samples was cloned into Escherichia coli and screened for expression of resistance to ampicillin or sulphonamide, two of the most common resistances found by array. The functional ampicillin resistance screen recovered genes encoding components of a predicted AcrRAB efflux pump. In the functional sulphonamide resistance screen, folP genes were recovered encoding mutant dihydropteroate synthase, the target of sulphonamide action. The genes recovered from the functional screens were from the chromosomes of commensal species that are opportunistically pathogenic and capable of exchanging DNA with related pathogenic species. Genes identified by microarray were not recovered in the activity-based screen, indicating that these two methods can be complementary in facilitating the identification of a range of resistance mechanisms present within the human microbiome. It also provides further evidence of the diverse reservoir of resistance mechanisms present in bacterial populations in the human gut and saliva. In future the methods described in this study can be used to monitor changes in the resistome in response to antibiotic therapy.

dc.format.extent0-0
dc.format.mediumElectronic-eCollection
dc.languageen
dc.language.isoen
dc.publisherPublic Library of Science (PLoS)
dc.subjectAmino Acid Sequence
dc.subjectAnti-Infective Agents
dc.subjectDihydropteroate Synthase
dc.subjectDrug Resistance, Microbial
dc.subjectFeces
dc.subjectHealthy Volunteers
dc.subjectHumans
dc.subjectMetagenome
dc.subjectMicroarray Analysis
dc.subjectMicrobial Sensitivity Tests
dc.subjectMicrobiota
dc.subjectMolecular Sequence Data
dc.subjectSaliva
dc.subjectSequence Alignment
dc.titleApplication of Microarray and Functional-Based Screening Methods for the Detection of Antimicrobial Resistance Genes in the Microbiomes of Healthy Humans
dc.typejournal-article
dc.typeJournal Article
dc.typeResearch Support, Non-U.S. Gov't
plymouth.author-urlhttps://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000330283100162&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=11bb513d99f797142bcfeffcc58ea008
plymouth.issue1
plymouth.volume9
plymouth.publication-statusPublished online
plymouth.journalPLoS ONE
dc.identifier.doi10.1371/journal.pone.0086428
plymouth.organisational-group/Plymouth
plymouth.organisational-group/Plymouth/Faculty of Health
plymouth.organisational-group/Plymouth/Faculty of Health/School of Biomedical Sciences
plymouth.organisational-group/Plymouth/REF 2021 Researchers by UoA
plymouth.organisational-group/Plymouth/REF 2021 Researchers by UoA/UoA01 Clinical Medicine
plymouth.organisational-group/Plymouth/Users by role
plymouth.organisational-group/Plymouth/Users by role/Academics
dc.publisher.placeUnited States
dcterms.dateAccepted2013-12-07
dc.identifier.eissn1932-6203
dc.rights.embargoperiodNot known
rioxxterms.versionofrecord10.1371/journal.pone.0086428
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.typeJournal Article/Review


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