Rapid identification and classification of Mycobacterium spp. using whole-cell protein barcodes with matrix assisted laser desorption ionization time of flight mass spectrometry in comparison with multigene phylogenetic analysis |
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Authors: | Jun Wang Wen Feng Chen Qing X. Li |
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Affiliation: | 1. Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu 96822, HI, USA;2. State Key Laboratories for Agro-Biotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China |
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Abstract: | The need of quick diagnostics and increasing number of bacterial species isolated necessitate development of a rapid and effective phenotypic identification method. Mass spectrometry (MS) profiling of whole cell proteins has potential to satisfy the requirements. The genus Mycobacterium contains more than 154 species that are taxonomically very close and require use of multiple genes including 16S rDNA for phylogenetic identification and classification. Six strains of five Mycobacterium species were selected as model bacteria in the present study because of their 16S rDNA similarity (98.4–99.8%) and the high similarity of the concatenated 16S rDNA, rpoB and hsp65 gene sequences (95.9–99.9%), requiring high identification resolution. The classification of the six strains by MALDI TOF MS protein barcodes was consistent with, but at much higher resolution than, that of the multi-locus sequence analysis of using 16S rDNA, rpoB and hsp65. The species were well differentiated using MALDI TOF MS and MALDI BioTyper™ software after quick preparation of whole-cell proteins. Several proteins were selected as diagnostic markers for species confirmation. An integration of MALDI TOF MS, MALDI BioTyper™ software and diagnostic protein fragments provides a robust phenotypic approach for bacterial identification and classification. |
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Keywords: | Matrix assisted laser desorption ionization time of flight Mycobacterium Bacterial classification Bacterial identification Bacterial barcode Whole-cell protein barcode |
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