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J Clin Microbiol. 2015 Aug;53(8):2632-40. doi: 10.1128/JCM.00813-15. Epub 2015 Jun 10.

Identification of Highly Pathogenic Microorganisms by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry: Results of an Interlaboratory Ring Trial.

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Robert Koch Institute, Proteomics and Spectroscopy (ZBS 6), Berlin, Germany.
Public Health Agency of Sweden, Solna, Sweden.
Austrian Agency for Health and Food Safety, Vienna, Austria.
National Center for Epidemiology, Department of Bacteriology, Budapest, Hungary.
Friedrich-Loeffler-Institut, Institute of Bacterial Infections and Zoonoses, Jena, Germany.
Bundeswehr Institute of Microbiology, Munich, Germany.
Norwegian Institute of Public Health, Oslo, Norway.
National Institute for Nuclear, Chemical and Biological Protection, Milin, Czech Republic.
National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
Spiez Laboratory, Federal Office for Civil Protection, Spiez, Switzerland.
Mabritec AG, Riehen, Switzerland.
Microbiology Laboratory and Infectious Diseases Biorepository, L. Spallanzani National Institute for Infectious Diseases, Rome, Italy.
Robert Koch Institute, Highly Pathogenic Microorganisms (ZBS 2), Berlin, Germany.
Robert Koch Institute, Highly Pathogenic Microorganisms (ZBS 2), Berlin, Germany


In the case of a release of highly pathogenic bacteria (HPB), there is an urgent need for rapid, accurate, and reliable diagnostics. MALDI-TOF mass spectrometry is a rapid, accurate, and relatively inexpensive technique that is becoming increasingly important in microbiological diagnostics to complement classical microbiology, PCR, and genotyping of HPB. In the present study, the results of a joint exercise with 11 partner institutions from nine European countries are presented. In this exercise, 10 distinct microbial samples, among them five HPB, Bacillus anthracis, Brucella canis, Burkholderia mallei, Burkholderia pseudomallei, and Yersinia pestis, were characterized under blinded conditions. Microbial strains were inactivated by high-dose gamma irradiation before shipment. Preparatory investigations ensured that this type of inactivation induced only subtle spectral changes with negligible influence on the quality of the diagnosis. Furthermore, pilot tests on nonpathogenic strains were systematically conducted to ensure the suitability of sample preparation and to optimize and standardize the workflow for microbial identification. The analysis of the microbial mass spectra was carried out by the individual laboratories on the basis of spectral libraries available on site. All mass spectra were also tested against an in-house HPB library at the Robert Koch Institute (RKI). The averaged identification accuracy was 77% in the first case and improved to >93% when the spectral diagnoses were obtained on the basis of the RKI library. The compilation of complete and comprehensive databases with spectra from a broad strain collection is therefore considered of paramount importance for accurate microbial identification.

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