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Advances in MALDI and Laser-Induced Soft Ionization Mass Spectrometry pp 197–225Cite as

MALDI Biotyping for Microorganism Identification in Clinical Microbiology

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Abstract

In 1996, independent publications demonstrated the application of MALDI-TOF MS for microorganism identification using whole-cell profile mass spectra. As these were characteristic for distinct bacterial and fungal species, they could be used as a molecular fingerprint. Subsequently, sample preparation, data evaluation with bioinformatics and further aspects of the technology were further improved. Ease-of-use, rapidity and accuracy of this technology finally led to the implementation of MALDI-TOF MS into routine analytical work, particularly in clinical microbiology.

Today, microorganism identification by MALDI-TOF MS is performed in many clinical microbiology laboratories and is increasingly replacing the conventional methods utilized for decades. In this chapter, the impact of this technology on clinical diagnostics is described. Furthermore, exemplary protocols for microorganism identification are presented. Specialized protocols even allow identification of more demanding microorganisms like mycobacteria and filamentous fungi as well as identification from liquid culture media inoculated with patient specimen. Some analytical systems allow the extension of reference databases with own entries and an insight into evaluation algorithms, interpretation of results and creation of own references is given.

Further, MALDI-TOF MS has proven its utility in identification of food-borne and veterinary microorganisms. These topics as well as regulatory aspects together with the necessary steps for qualification and validation are also covered for the interested reader.

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Notes

  1. 1.

    The slightly rougher structure of ground steel targets can facilitate the smearing to the right thickness.

  2. 2.

    Most culture media are blood-based. The haemoglobin in these media can produce artificial peaks disturbing the spectral acquisition or deteriorating the identification result. Thus, accidental transfers of parts of the culture media to the target should be avoided.

  3. 3.

    For blood culture bottles that contain charcoal for adsorbing antimicrobial substances, filtering the blood culture/lysis buffer mixture in the first steps with a spin column like SigmaPrep (cat. no. SC1000-1KT, Sigma-Aldrich; centrifugation for 2 min at 2000 rpm) can remove the distracting particles. For blood cultures with resin particles this procedure is not needed.

  4. 4.

    Sometimes it is advisable to accept slightly lower score values. In the rare cases when more than one pathogenic microorganism is suspected in the blood culture or when Gram staining already shows multiple morphologies in microscopy, mixture algorithms can be applied for identification. However, in general, blood cultures only contain one microorganism.

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Authors and Affiliations

  1. Department of Medical Microbiology, MVZ Dr. Eberhard & Partner Dortmund (ÜBAG), Balkenstr. 17-19, 44137, Dortmund, Germany

    Arthur B. Pranada

  2. Elisabeth-Segelken-Str. 7, 28357, Bremen, Germany

    Gerold Schwarz

  3. Bruker Daltonik GmbH, Fahrenheitstr. 4, 28359, Bremen, Germany

    Markus Kostrzewa

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  1. Arthur B. Pranada
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  2. Gerold Schwarz
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  3. Markus Kostrzewa
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Correspondence to Arthur B. Pranada .

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  1. Department of Chemistry, University of Reading, Reading, Berkshire, United Kingdom

    Rainer Cramer

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Pranada, A.B., Schwarz, G., Kostrzewa, M. (2016). MALDI Biotyping for Microorganism Identification in Clinical Microbiology. In: Cramer, R. (eds) Advances in MALDI and Laser-Induced Soft Ionization Mass Spectrometry. Springer, Cham. https://doi.org/10.1007/978-3-319-04819-2_11

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