title: Rapid identification of bacteria and yeasts from positive-blood-culture bottles by using a lysis-filtration method and matrix-assisted laser desorption ionization-time of flight mass spectrum analysis with the SARAMIS database.
abstract: Rapid identification of microorganisms causing bloodstream infections directly from a positive blood culture would decrease the time to directed antimicrobial therapy and greatly improve patient care. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a fast and reliable method for identifying microorganisms from positive culture. This study evaluates the performance of a novel filtration-based method for processing positive-blood-culture broth for immediate identification of microorganisms by MALDI-TOF with a Vitek MS research-use-only system (VMS). BacT/Alert non-charcoal-based blood culture bottles that were flagged positive by the BacT/Alert 3D system were included. An aliquot of positive-blood-culture broth was incubated with lysis buffer for 2 to 4 min at room temperature, the resulting lysate was filtered through a membrane, and harvested microorganisms were identified by VMS. Of the 259 bottles included in the study, VMS identified the organisms in 189 (73%) cultures to the species level and 51 (19.7%) gave no identification (ID), while 6 (2.3%) gave identifications that were considered incorrect. Among 131 monomicrobic isolates from positive-blood-culture bottles with one spot having a score of 99.9%, the IDs for 131 (100%) were correct to the species level. In 202 bottles where VMS was able to generate an ID, the IDs for 189 (93.6%) were correct to the species level, whereas the IDs provided for 7 isolates (3.5%) were incorrect. In conclusion, this method does not require centrifugation and produces a clean spectrum for VMS analysis in less than 15 min. This study demonstrates the effectiveness of the new lysis-filtration method for identifying microorganisms directly from positive-blood-culture bottles in a clinical setting.
authors.affiliation: Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Fothergill
authors.forename: Vyjayanti
authors.initials: V
authors.lastname: Kasinathan
authors.forename: Jay
authors.initials: J
authors.lastname: Hyman
authors.forename: John
authors.initials: J
authors.lastname: Walsh
authors.forename: Tim
authors.initials: T
authors.lastname: Drake
authors.forename: Yun F Wayne
authors.initials: YF
authors.lastname: Wang
mesh_terms: D001419:Bacteria / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D001769:Blood / Q000382:microbiology*; D005374:Filtration / Q000379:methods; D005658:Fungi / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D018805:Sepsis / Q000175:diagnosis*; D013048:Specimen Handling / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors