Identification of Biomarkers of Whole Coxiella burnetii Phase I by MALDI-TOF Mass Spectrometry

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) specific biomarkers have been shown to be an effective tool for identifying microorganisms. In this study, we demonstrate the feasibility of using this technique to detect the obligate intracellular bacterium...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2004-07, Vol.76 (14), p.4017-4022
Main Authors: SHAW, Edward I., MOURA, Hercules, WOOLFITT, Adrian R., OSPINA, Maria, THOMPSON, Herbert A., BARR, John R.
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Bacterial Proteins - analysis
Biomarkers - analysis
Chemistry
Chick Embryo
Coxiella burnetii - isolation & purification
Eggs - microbiology
Exact sciences and technology
Spectrometric and optical methods
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
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Summary:Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) specific biomarkers have been shown to be an effective tool for identifying microorganisms. In this study, we demonstrate the feasibility of using this technique to detect the obligate intracellular bacterium Coxiella burnetii, a category B bioterrorism agent. Specific biomarkers were detected in C. burnetii Nine Mile phase I (NMI) strain purified from embryonated egg yolk sac preparations. Whole organisms were applied directly to the MALDI target. MALDI-TOF MS analysis of C. burnetii NMI grown and purified at different times and places revealed a group of unique, characteristic, and reproducible spectral markers in the mass range of 1000-25000 Da. Statistical analysis of the averaged centroided masses uncovered at least 24 peptides or biomarkers. Three biomarkers observed in the MALDI-TOF MS spectrum consistently matched proteins that had been previously described in C. burnetii, one of them being the small cell variant protein A. MALDI-TOF MS analysis of whole organisms represents a sensitive and specific option for characterizing C. burnetii isolates, especially when coupled with antigen capture techniques. The method also has potential for several applications in basic microbial research, including regulation of gene expression.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac030364k