Use of MALDI-TOF MS for Identification of Nontuberculous Mycobacterium Species Isolated from Clinical Specimens

The aim of this study was to compare the results obtained for identification by MALDI-TOF of nontuberculous mycobacteria (NTM) isolated in clinical samples with those obtained by GenoType Mycobacterium CM/AS (common mycobacteria/additional species). A total of 66 Mycobacterium isolates from various clinical specimens (mainly respiratory) were tested in this study. They were identified using MALDI-TOF Bruker from strains isolated in Lowenstein, following the recommended protocol of heat inactivation and extraction, and were simultaneously analyzed through hybridization by GenoType Mycobacterium from liquid culture MGIT. Our results showed that identification by MALDI-TOF was correct in 98.4% (65/66) of NTM isolated in our clinical practice (M. avium, M. intracellulare, M. abscessus, M. chelonae, M. fortuitum, M. mucogenicum, M. kansasii, and M. scrofulaceum). MALDI-TOF was found to be an accurate, rapid, and cost-effective system for identification of mycobacteria species.

[1]  A. Steigerwalt,et al.  Clinical and Laboratory Features of Mycobacterium porcinum , 2004, Journal of Clinical Microbiology.

[2]  Robert Horsburgh,et al.  An official ATS/IDSA statement: diagnosis, treatment, and prevention of nontuberculous mycobacterial diseases. , 2007, American journal of respiratory and critical care medicine.

[3]  Markus Kostrzewa,et al.  Comparison of MALDI-TOF MS with HPLC and nucleic acid sequencing for the identification of Mycobacterium species in cultures using solid medium and broth. , 2014, American journal of clinical pathology.

[4]  S. Rüsch-Gerdes,et al.  Evaluation of the GenoType Mycobacterium Assay for Identification of Mycobacterial Species from Cultures , 2006, Journal of Clinical Microbiology.

[5]  Linda S. Guthertz,et al.  Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification of Mycobacterium Species , 2001, Clinical Microbiology Reviews.

[6]  S. Butler-Wu,et al.  Comparison of the Bruker Biotyper and Vitek MS Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry Systems for Identification of Mycobacteria Using Simplified Protein Extraction Protocols , 2013, Journal of Clinical Microbiology.

[7]  U. Singh,et al.  Rapid identification of clinical mycobacterial isolates by protein profiling using matrix assisted laser desorption ionization-time of flight mass spectrometry. , 2013, Indian journal of medical microbiology.

[8]  E. Tortoli Clinical manifestations of nontuberculous mycobacteria infections. , 2009, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[9]  V. Rotimi,et al.  Comparative evaluation of two matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) systems for the identification of clinically significant yeasts. , 2014, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[10]  E. Mcgrath,et al.  Increased prevalence of non-tuberculous mycobacteria infection , 2007, The Lancet.

[11]  M. Slaný,et al.  The influence of culture conditions on the identification of Mycobacterium species by MALDI-TOF MS profiling. , 2014, FEMS microbiology letters.

[12]  P. Murray,et al.  Identification of Mycobacteria in Solid-Culture Media by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry , 2011, Journal of Clinical Microbiology.

[13]  U. Göbel,et al.  Occurrence and Clinical Relevance of Mycobacterium chimaera sp. nov., Germany , 2008, Emerging infectious diseases.

[14]  Y. Wang,et al.  Comparison of Heat Inactivation and Cell Disruption Protocols for Identification of Mycobacteria from Solid Culture Media by Use of Vitek Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry , 2013, Journal of Clinical Microbiology.

[15]  P. Berche,et al.  Rapid Identification of Mycobacterial Whole Cells in Solid and Liquid Culture Media by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry , 2010, Journal of Clinical Microbiology.

[16]  M. Almuzara,et al.  Comparison of the Bruker MALDI-TOF Mass Spectrometry System and Conventional Phenotypic Methods for Identification of Gram-Positive Rods , 2014, PloS one.

[17]  R. Kroppenstedt,et al.  Proposal to elevate the genetic variant MAC-A, included in the Mycobacterium avium complex, to species rank as Mycobacterium chimaera sp. nov. , 2004, International journal of systematic and evolutionary microbiology.

[18]  Brockhoff Ej,et al.  Pulmonary Disease Associated with Nontuberculous Mycobacteria , 2011 .

[19]  Paulette Legarraga,et al.  [Impact of mass spectrometry by MALDI-TOF MS for the rapid identification of aerobic and anaerobic bacteria of clinical importance]. , 2013, Revista chilena de infectologia : organo oficial de la Sociedad Chilena de Infectologia.

[20]  E. Tortoli Impact of Genotypic Studies on Mycobacterial Taxonomy: the New Mycobacteria of the 1990s , 2003, Clinical Microbiology Reviews.

[21]  C. TamaraLeiva Evaluación de una técnica de hibridación reversa para identificación rápida de micobacterias en Chile , 2012 .

[22]  Frank,et al.  Rapid identification of mycobacteria to the species level by polymerase chain reaction and restriction enzyme analysis , 1993, Journal of clinical microbiology.

[23]  M. Wei,et al.  Nontuberculous Mycobacteria Infections in Immunocompromised Patients: Single Institution Experience , 2009, Journal of pediatric hematology/oncology.

[24]  K. Winthrop,et al.  Pulmonary Disease Associated with Nontuberculous Mycobacteria, Oregon, USA , 2011, Emerging infectious diseases.

[25]  M. Raponi,et al.  Advancement in the routine identification of anaerobic bacteria by MALDI-TOF mass spectrometry , 2013, European Journal of Clinical Microbiology & Infectious Diseases.

[26]  A. Rodloff,et al.  Discrimination of Enterobacteriaceae and Non-fermenting Gram Negative Bacilli by MALDI-TOF Mass Spectrometry , 2013, The open microbiology journal.

[27]  T. Adékambi Mycobacterium mucogenicum group infections: a review. , 2009, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[28]  Didier Raoult,et al.  rpoB gene sequence-based characterization of emerging non-tuberculous mycobacteria with descriptions of Mycobacterium bolletii sp. nov., Mycobacterium phocaicum sp. nov. and Mycobacterium aubagnense sp. nov. , 2006, International journal of systematic and evolutionary microbiology.

[29]  P. García-Martos,et al.  Infecciones por micobacterias de crecimiento rápido , 2012 .

[30]  Laura Ferreira,et al.  Aplicaciones de la proteómica en el laboratorio de Microbiología Clínica , 2012 .