title: Rapid identification of bacteria from bioMérieux BacT/ALERT blood culture bottles by MALDI-TOF MS.
abstract: Several studies have reported poor results when trying to identify microorganisms directly from the bioMérieux BacT/ALERT blood culture system using matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry. The aim of this study is to evaluate two new methods, Sepsityper and an enrichment method for direct identification of microorganisms from this system. For both methods the samples were processed using the Bruker Microflex LT mass spectrometer (Biotyper) using the Microflex Control software to obtain spectra. The results from direct analysis were compared with those obtained by subculture and subsequent identification. A total of 350 positive blood cultures were processed simultaneously by the two methods. Fifty-three cultures were polymocrobial or failed to grow any organism on subculture, and these results were not included as there was either no subculture result, or for polymicrobial cultures it was known that the Biotyper would not be able to distinguish the constituent organisms correctly. Overall, the results showed that, contrary to previous reports, it is possible to identify bacteria directly from bioMérieux blood culture bottles, as 219/297 (74%) correct identifications were obtained using the Bruker Sepsityper method and 228/297 (77%) were obtained for the enrichment method when there is only one organism was present. Although the enrichment method was simpler, the reagent costs for the Sepsityper method were approximately pound 4.00 per sample compared to pound 0.50. An even simpler and cheaper method, which was less labour-intensive and did not require further reagents, was investigated. Seventy-seven specimens from positive signalled blood cultures were analysed by inoculating prewarmed blood agar plates and analysing any growth after 1-, 2- and 4-h periods of incubation at 37 degrees C, by either direct transfer or alcohol extraction. This method gave the highest number of correct identifications, 66/77 (86%), and was cheaper and less labour-intensive than either of the two above methods.
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: J D
authors.initials: JD
authors.lastname: Haigh
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: I M
authors.initials: IM
authors.lastname: Green
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: D
authors.initials: D
authors.lastname: Ball
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: M
authors.initials: M
authors.lastname: Eydmann
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: M
authors.initials: M
authors.lastname: Millar
authors.affiliation: Microbiology, The Royal London Hospital, 3rd Floor Pathology and Pharmacy Building, 80 Newark Street, London E1 2ES.
authors.forename: M
authors.initials: M
authors.lastname: Wilks
mesh_terms: D001419:Bacteria / Q000145:classification / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques / Q000295:instrumentation* / Q000379:methods*; D001769:Blood / Q000382:microbiology*; D003470:Culture Media; D006801:Humans; D015203:Reproducibility of Results; D012984:Software; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013210:Staphylococcus / Q000145:classification / Q000302:isolation & purification; D013997:Time Factors
chemical_list: D003470:Culture Media