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Journal of Clinical Microbiology, June 2008, p. 1946-1954, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.00157-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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{triangledown}

A. Mellmann,1* J. Cloud,2 T. Maier,3 U. Keckevoet,1 I. Ramminger,1 P. Iwen,4 J. Dunn,5 G. Hall,6 D. Wilson,6 P. LaSala,7 M. Kostrzewa,3 and D. Harmsen8

Institute for Hygiene, University Hospital Muenster, Muenster D-48149, Germany,1 ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah,2 Bruker Daltonik GmbH, Leipzig, Germany,3 Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska,4 Cook Children's Medical Center, Fort Worth, Texas,5 Microbiology, Cleveland Clinic Foundation, Cleveland, Ohio,6 Clinical Microbiology, Department of Pathology, University of Texas Medical Branch, Galveston, Texas,7 Department for Periodontology, University Hospital Muenster, Muenster D-48149, Germany8

Received 25 January 2008/ Returned for modification 21 March 2008/ Accepted 1 April 2008

Nonfermenting bacteria are ubiquitous environmental opportunists that cause infections in humans, especially compromised patients. Due to their limited biochemical reactivity and different morphotypes, misidentification by classical phenotypic means occurs frequently. Therefore, we evaluated the use of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for species identification. By using 248 nonfermenting culture collection strains composed of 37 genera most relevant to human infections, a reference database was established for MALDI-TOF MS-based species identification according to the manufacturer's recommendations for microflex measurement and MALDI BioTyper software (Bruker Daltonik GmbH, Leipzig, Germany), i.e., by using a mass range of 2,000 to 20,000 Da and a new pattern-matching algorithm. To evaluate the database, 80 blind-coded clinical nonfermenting bacterial strains were analyzed. As a reference method for species designation, partial 16S rRNA gene sequencing was applied. By 16S rRNA gene sequencing, 57 of the 80 isolates produced a unique species identification (≥99% sequence similarity); 11 further isolates gave ambiguous results at this threshold and were rated as identified to the genus level only. Ten isolates were identified to the genus level (≥97% similarity); and two isolates had similarity values below this threshold, were counted as not identified, and were excluded from further analysis. MALDI-TOF MS identified 67 of the 78 isolates (85.9%) included, in agreement with the results of the reference method; 9 were misidentified and 2 were unidentified. The identities of 10 randomly selected strains were 100% correct when three different mass spectrometers and four different cultivation media were used. Thus, MALDI-TOF MS-based species identification of nonfermenting bacteria provided accurate and reproducible results within 10 min without any substantial costs for consumables.

* Corresponding author. Mailing address: Institut fuer Hygiene, Universitätsklinikum Muenster, Robert-Koch-Str. 41, Muenster D-48149, Germany. Phone: 49 251 8352316. Fax: 49 251 8355688. E-mail: mellmann{at}uni-muenster.de

{triangledown} Published ahead of print on 9 April 2008.

Journal of Clinical Microbiology, June 2008, p. 1946-1954, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.00157-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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