title: Development of a rapid diagnostic assay based on magnetic bead purification of OXA-β-lactamase mRNA.
abstract: To countermeasure the global spread of β-lactamases, we developed a rapid molecular test for the highly variable OXA-β-lactamases that allows minimizing the time to effective treatment.
  
  
  
  OXA-mRNA was specifically enriched from total RNA using group-specific biotinylated DNA probes and streptavidin-coated magnetic beads. Phylogenetic OXA groups were distinguished by PCR product size.
  
  
  
  This mRNA fishing method is highly sensitive, yielding specific results from as little as 1 ng total RNA. It enables discrimination of OXA-extended substrate spectrum β-lactamases and carbapenemases and the semi-quantitative detection of highly expressed ISAba1-controlled variants.
  
  
  
  Targeting mRNA with specific probes on magnetic beads will allow for adaptation to automated systems, such as point-of-care diagnostics.
authors.affiliation: Center for Infectious Diseases & Infection Control, Jena University Hospital, Jena, Germany.|Infectognostics Research Campus, Jena, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Brandt
authors.affiliation: Institute for Medical Microbiology, Immunology & Hygiene, University Hospital of Cologne, Cologne, Germany.
authors.forename: Esther
authors.initials: E
authors.lastname: Zander
authors.affiliation: Nosocomial Pathogens & Antibiotic Resistance, Robert Koch Institute, Wernigerode, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Pfeifer
authors.affiliation: Infectognostics Research Campus, Jena, Germany.|Alere Technologies GmbH, Jena, Germany.
authors.forename: Sascha D
authors.initials: SD
authors.lastname: Braun
authors.affiliation: Infectognostics Research Campus, Jena, Germany.|Alere Technologies GmbH, Jena, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Ehricht
authors.affiliation: Center for Infectious Diseases & Infection Control, Jena University Hospital, Jena, Germany.|Infectognostics Research Campus, Jena, Germany.
authors.forename: Oliwia
authors.initials: O
authors.lastname: Makarewicz
authors.affiliation: Center for Infectious Diseases & Infection Control, Jena University Hospital, Jena, Germany.|Infectognostics Research Campus, Jena, Germany.
authors.forename: Mathias W
authors.initials: MW
authors.lastname: Pletz
mesh_terms: D000151:Acinetobacter Infections / Q000175:diagnosis* / Q000382:microbiology; D040981:Acinetobacter baumannii / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D004755:Enterobacteriaceae / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D004756:Enterobacteriaceae Infections / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D008280:Magnetics / Q000295:instrumentation / Q000379:methods*; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D001618:beta-Lactamases
keywords: Acinetobacter baumannii; CHDL; Dynabeads; ESBL; carbapenem resistance; multiplex; sequence homology