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2013-11-05Zeitschriftenartikel DOI: 10.1371/journal.pone.0080079
Direct RNA-Based Detection and Differentiation of CTX-M-Type Extended-Spectrum β-Lactamases (ESBL)
dc.contributor.authorStein, Claudia
dc.contributor.authorMakarewicz, Oliwia
dc.contributor.authorPfeifer, Yvonne
dc.contributor.authorBrandt, Christian
dc.contributor.authorRamos, João Costa
dc.contributor.authorKlinger, Mareike
dc.contributor.authorPletz, Mathias W.
dc.date.accessioned2018-05-07T17:16:01Z
dc.date.available2018-05-07T17:16:01Z
dc.date.created2013-12-12
dc.date.issued2013-11-05none
dc.identifier.otherhttp://edoc.rki.de/oa/articles/reps3LTmJZsGo/PDF/27ctDoYusMTpY.pdf
dc.identifier.urihttp://edoc.rki.de/176904/1745
dc.description.abstractThe current global spread of multi-resistant Gram-negatives, particularly extended spectrum β-lactamases expressing bacteria, increases the likelihood of inappropriate empiric treatment of critically ill patients with subsequently increased mortality. From a clinical perspective, fast detection of resistant pathogens would allow a pre-emptive correction of an initially inappropriate treatment. Here we present diagnostic amplification-sequencing approach as proof of principal based on the fast molecular detection and correct discrimination of CTX-M-β-lactamases, the most frequent ESBL family. The workflow consists of the isolation of total mRNA and CTX-M-specific reverse transcription (RT), amplification and pyrosequencing. Due to the high variability of the CTX-M-β-lactamase-genes, degenerated primers for RT, qRT as well as for pyrosequencing, were used and the suitability and discriminatory performance of two conserved positions within the CTX-M genes were analyzed, using one protocol for all isolates and positions, respectively. Using this approach, no information regarding the expected CTX-M variant is needed since all sequences are covered by these degenerated primers. The presented workflow can be conducted within eight hours and has the potential to be expanded to other β-lactamase families.eng
dc.language.isoeng
dc.publisherRobert Koch-Institut, Infektionskrankheiten / Erreger
dc.subjectbeta-Lactamases/geneticseng
dc.subjectReal-Time Polymerase Chain Reactioneng
dc.subjectMutationeng
dc.subjectEscherichia coli/enzymologyeng
dc.subjectRNA/geneticseng
dc.subjectReverse Transcriptioneng
dc.subject.ddc610 Medizin
dc.titleDirect RNA-Based Detection and Differentiation of CTX-M-Type Extended-Spectrum β-Lactamases (ESBL)
dc.typeperiodicalPart
dc.identifier.urnurn:nbn:de:0257-10033983
dc.identifier.doi10.1371/journal.pone.0080079
dc.identifier.doihttp://dx.doi.org/10.25646/1670
local.edoc.container-titlePLoS ONE
local.edoc.container-textStein C, Makarewicz O, Pfeifer Y, Brandt C, Ramos JC, et al. (2013) Direct RNA-Based Detection and Differentiation of CTX-M-Type Extended-Spectrum β-Lactamases (ESBL). PLoS ONE 8(11): e80079.
local.edoc.fp-subtypeArtikel
local.edoc.type-nameZeitschriftenartikel
local.edoc.container-typeperiodical
local.edoc.container-type-nameZeitschrift
local.edoc.container-urlhttp://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0080079
local.edoc.container-publisher-namePublic Library of Science
local.edoc.container-volume8
local.edoc.container-issue11
local.edoc.container-year2013

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