A metabolomics approach to characterise and identify various Mycobacterium species.
We investigated the potential use of gas chromatography mass spectrometry (GC-MS), in combination with multivariate statistical data processing, to build a model for the classification of various tuberculosis (TB) causing, and non-TB Mycobacterium species, on the basis of their characteristic metabolite profiles. A modified Bligh-Dyer extraction procedure was used to extract lipid components from Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium bovis, and Mycobacterium kansasii cultures. Principle component analyses (PCA) of the GC-MS generated data showed a clear differentiation between all the Mycobacterium species tested. Subsequently, the 12 compounds best describing the variation between the sample groups were identified as potential metabolite markers, using PCA and partial least-squares discriminant analysis (PLS-DA). These metabolite markers were then used to build a discriminant classification model based on Bayes' theorem, in conjunction with multivariate kernel density estimation. This model subsequently correctly classified 2 "unknown" samples for each of the Mycobacterium species analysed, with probabilities ranging from 72 to 100%. Furthermore, Mycobacterium species classification could be achieved in less than 16 h, and the detection limit for this approach was 1×10(3)bacteriamL(-1). This study proves the capacity of a GC-MS, metabolomics pattern recognition approach for its possible use in TB diagnostics and disease characterisation.
School for Physical and Chemical Sciences, Centre for Human Metabonomics, North-West University, Potchefstroom, 2520, South Africa.
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Olivier
Du Toit
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Loots
D001431:Bacteriological Techniques / Q000379:methods*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D055432:Metabolomics / Q000379:methods*; D015233:Models, Statistical; D009161:Mycobacterium / Q000737:chemistry* / Q000145:classification* / Q000302:isolation & purification / Q000378:metabolism; D009164:Mycobacterium Infections / Q000175:diagnosis* / Q000382:microbiology*