Species identification of Campylobacter jejuni ssp. jejuni and C. coli by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and PCR

Abstract

The Campylobacter species strains (n = 42; isolated from clinical samples and deposited in Czech National Collection of Type Cultures, Prague) originally phenotypically (and biochemically) identified as Campylobacter jejuni were re-classified using molecular biological and mass spectrometric methods. Whole-cell MALDI-TOF MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry) separated the isolates into two genetically related strains--C. jejuni (n = 26) and C. coli (n = 16) and, moreover, distinguished the intimate details in the group of tested strains. It also made it possible to create the MALDI-TOF MS dendrogram; similarly, the spectral characteristics were used for the 3D cluster analysis. Polymerase chain reaction (PCR) confirmed the results obtained by mass spectrometry. Both methods (PCR and MALDI-TOF MS) gave the same results which supports their suitability in the rapid and accurate Campylobacter-species determination.

Matrix-Assisted Laser Desorption Ionization (MALDI)-Time of Flight Mass Spectrometry- and MALDI Biotyper-Based Identification of Cultured Biphenyl-Metabolizing Bacteria from Contaminated Horseradish Rhizosphere Soil

“…It has been proved that once the samples for analyses have been treated in the same way, the generated spectra are quite reproducible and unique fingerprints are achieved with different strains, as was shown for E. coli (3), pathogenic Neisseria strains (26), staphylococci (15), listeriae (6), campylobacters (34), mycobacteria (23), and others. To the best of our knowledge, this study is the first one to apply MALDI-TOF MS to environmental isolates (which are potentially relevant for bioremediation).…”

Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: a Fundamental Shift in the Routine Practice of Clinical Microbiology

“…The study determined that MALDI-TOF MS could be used to differentiate between the two genera as well as related members of different species through the identification of specific spectral peaks corresponding to biomarkers (228). Subsequent studies (229,230) further examined the ability of MALDI-TOF MS to provide rapid and accurate species-level identifications for members of the genus, with encouraging results.…”

Mass Spectrometric Identification of <i>Propionibacterium</i> Isolates Requires Database Enrichment

“…All strains were grown on Columbia blood agar containing 5% whole sheep blood (BioMerieux, Marcy l'étoile, France) at 37˚C in anaerobic atmosphere. In addition, in order to estimate the validity of MALDI-TOF MS for P. acnes identification, the main pathogenic Propionibacterium species, 48 additional P. acnes isolates from clinical samples were tested by MS. P. acnes isolates were cultivated on Columbia blood agar at 37˚C in anaerobic atmosphere as described above from blood (12), cardiac valve biopsies (2), surgically-removed pace-makers (3), a lung biopsy (1), lymph node biopsies (3), bone biopsies (3), cerebrospinal fluid (1), skin biopsy samples (23). The identification of these strains was confirmed by PCR amplification and sequencing of the 16S RNA, as previously described [43].…”

“…MALDI-TOF MS was applied with success to identify Arthrobacter species [19], Bacteroides sp. [20], Bartonella [21], Burkholderia cepacia [22], Campylobacter species [23], Clostridium species [24], Coxiella burnetii [25], Erwinia species [26], Escherichia coli [27], Francisella tularensis [28], Helicobacter pylori [29], Legionella species [30], Listeria species [31], Neisseria species [32], Salmonella species [33], staphylococci [34], Vibrio species [35], viridans streptococci [36], Yersinia enterocolitica [27], nonfermenting bacteria [37,38], and oral anaerobic bacteria [39]. Moreover, recent studies emphasized that MALDI-TOF MS identification was an efficient and cost-effective method for rapid and routine identification of bacterial isolates [17,[40][41][42].…”