title: Rapid inactivation of Mycobacterium and nocardia species before identification using matrix-assisted laser desorption ionization-time of flight mass spectrometry.
abstract: The identification of mycobacteria outside biocontainment facilities requires that the organisms first be rendered inactive. Exposure to 70% ethanol (EtOH) either before or after mechanical disruption was evaluated in order to establish a safe, effective, and rapid inactivation protocol that is compatible with identification of Mycobacterium and Nocardia species using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). A combination of 5 min of bead beating in 70% EtOH followed by a 10-min room temperature incubation period was found to be rapidly bactericidal and provided high-quality spectra compared to spectra obtained directly from growth on solid media. The age of the culture, the stability of the refrigerated or frozen lysates, and freeze-thaw cycles did not adversely impact the quality of the spectra or the identification obtained.
authors: bioMérieux, Inc., Durham, North Carolina, USA william.dunne@biomerieux.com.
authors: W Michael
authors: WM
authors: Dunne
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Kirk
authors: K
authors: Doing
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Elizabeth
authors: E
authors: Miller
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Eric
authors: E
authors: Miller
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Erik
authors: E
authors: Moreno
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Mehdi
authors: M
authors: Baghli
authors: bioMérieux, La Balme Les Grottes, France.
authors: Sandrine
authors: S
authors: Mailler
authors: bioMérieux, La Balme Les Grottes, France.
authors: Victoria
authors: V
authors: Girard
authors: bioMérieux, La Balme Les Grottes, France.
authors: Alex
authors: A
authors: van Belkum
authors: bioMérieux, Inc., Durham, North Carolina, USA.
authors: Parampal
authors: P
authors: Deol
mesh_terms: D004203:Disinfection / Q000379:methods*; D000431:Ethanol / Q000633:toxicity; D006801:Humans; D055595:Mechanical Phenomena; D050296:Microbial Viability / Q000187:drug effects; D009161:Mycobacterium / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology*; D009164:Mycobacterium Infections / Q000175:diagnosis; D009615:Nocardia / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology*; D009617:Nocardia Infections / Q000175:diagnosis; D013048:Specimen Handling / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013997:Time Factors
chemical_list: D000431:Ethanol