Direct Identification of Urinary Tract Pathogens from Urine Samples by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

ABSTRACT Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been suggested as a reliable method for bacterial identification from cultures. Direct analysis of clinical samples might increase the usefulness of this method, shortening the time for microorganism identification. We compared conventional methods for the diagnosis of urinary tract infections (UTIs) and identification of the urinary tract pathogens (automated screening, plate cultures, and identification based on biochemical characteristics) and a fast method based on conventional screening and MALDI-TOF MS. For this latter method, 4 ml of urine was centrifuged at a low-revolution setting (2,000 × g) to remove leukocytes and then at high revolutions (15,500 × g) to collect bacteria. The pellet was washed and then applied directly to the MALDI-TOF MS plate. Two hundred sixty urine samples, detected as positive by the screening device (UF-1000i), were processed by culture and MALDI-TOF MS. Twenty samples were positive in the screening device but negative in culture, and all of them were also negative by MALDI-TOF MS. Two-hundred thirty-five samples displayed significant growth of a single morphological type in culture. Two-hundred twenty of them showed bacterial growth of >105 CFU/ml. Microorganism identifications in this group were coincident at the species level in 202 cases (91.8%) and at the genus level in 204 cases (92.7%). The most frequent microorganism was Escherichia coli (173 isolates). MALDI-TOF MS identified this microorganism directly from the urine sample in 163 cases (94.2%). Our results show that MALDI-TOF MS allows bacterial identification directly from infected urine in a short time, with high accuracy, and especially when Gram-negative bacteria with high bacterial counts are involved.

[1]  T. Hooton,et al.  Diagnosis and treatment of uncomplicated urinary tract infection. , 1997, Infectious disease clinics of North America.

[2]  J. Sobel,et al.  Urinary tract infection: self-reported incidence and associated costs. , 2000, Annals of epidemiology.

[3]  P. Demirev,et al.  Characterization of intact microorganisms by MALDI mass spectrometry. , 2001, Mass spectrometry reviews.

[4]  Lex M Bouter,et al.  The urine dipstick test useful to rule out infections. A meta-analysis of the accuracy , 2004, BMC urology.

[5]  Trinad Chakraborty,et al.  Rapid Identification and Typing of Listeria Species by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry , 2008, Applied and Environmental Microbiology.

[6]  M. Erhard,et al.  Rapid Classification and Identification of Salmonellae at the Species and Subspecies Levels by Whole-Cell Matrix-Assisted Laser Desorption Ionization – Time of Flight Mass Spectrometry † , 2008 .

[7]  Markus Kostrzewa,et al.  Challenging the problem of clostridial identification with matrix-assisted laser desorption and ionization-time-of-flight mass spectrometry (MALDI-TOF MS). , 2008, Anaerobe.

[8]  A. Mellmann,et al.  Evaluation of Matrix-Assisted Laser Desorption Ionization-Time-of-Flight Mass Spectrometry in Comparison to 16S rRNA Gene Sequencing for Species Identification of Nonfermenting Bacteria , 2008, Journal of Clinical Microbiology.

[9]  P. Berche,et al.  Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Identification of Nonfermenting Gram-Negative Bacilli Isolated from Cystic Fibrosis Patients , 2008, Journal of Clinical Microbiology.

[10]  D. Raoult,et al.  Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. , 2009, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[11]  A. Hoerauf,et al.  Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry for Fast and Reliable Identification of Clinical Yeast Isolates , 2009, Journal of Clinical Microbiology.

[12]  T. Maier,et al.  Species identification of clinical isolates of Bacteroides by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry. , 2009, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[13]  R. Courcol,et al.  Identification bactérienne par spectrométrie de masse de type MALDI-TOF : évaluation au CHU de Lille , 2010 .

[14]  E. Urbán,et al.  ESCMID Study group on Antimicrobial resistance in anaerobic bacteria , 2011 .