Identification of Brucella spp. isolates and discrimination from the vaccine strain Rev.1 by MALDI-TOF mass spectrometry

Christoforidou, Sofia; Kyritsi, Maria; Boukouvala, Evridiki; Ekateriniadou, Loukia; Ζdragas, Αntonios; Samouris, Georgios; Hadjichristodoulou, Christos

doi:10.1016/j.mcp.2020.101533

Cited by 7 publications

(14 citation statements)

References 21 publications

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“…Initially, a cluster analysis on a subset of 17 field strains that represented the three B. melitensis biovars against three commercial B. melitensis Rev.1 strains had been performed. Two discriminating peaks were described in this study: Peak m/z 3,528 could be detected in all tested Greek B. melitensis field isolates but not in B. melitensis Rev.1, whereas peak m/z 7,328 was unique for the vaccine strain (Christoforidou et al, 2020). Strikingly, these biomarker peaks were not identified in our study, thus we compared the data published by Christoforidou et al (2020) with our data.…”

Section: Differentiation Of Brucella Melitensismentioning

confidence: 66%

“…Two discriminating peaks were described in this study: Peak m/z 3,528 could be detected in all tested Greek B. melitensis field isolates but not in B. melitensis Rev.1, whereas peak m/z 7,328 was unique for the vaccine strain (Christoforidou et al, 2020). Strikingly, these biomarker peaks were not identified in our study, thus we compared the data published by Christoforidou et al (2020) with our data. The first described biomarker with m/z 3,528, supposed to be only present in the spectra of B. melitensis Greek field isolates, could be found in the spectra of all Israeli field isolates, but also in the B. melitensis 16M reference strain and the Rev.1 strain (Figure 4A).…”

Section: Differentiation Of Brucella Melitensismentioning

confidence: 66%

“…The first described biomarker with m/z 3,528, supposed to be only present in the spectra of B. melitensis Greek field isolates, could be found in the spectra of all Israeli field isolates, but also in the B. melitensis 16M reference strain and the Rev.1 strain (Figure 4A). The second biomarker with m/z 7,328 neither occurred in the Greek B. melitensis field isolates (Christoforidou et al, 2020) nor in the B. melitensis field isolates from Israel (Figure 4A), but in the B. melitensis Rev.1 strain of both studies. However, this peak does not reflect a true biomarker for B. melitensis Rev.1 against the closely related B. melitensis 16M from the same MLVA group "melitensis Americas, " which also harbors this causative protein (Figure 4A).…”

Section: Differentiation Of Brucella Melitensismentioning

confidence: 91%

“…Based on our peak observations, we determined mass windows with a decision boundary to distinguish between ribosomal protein L24 and S12 variants, and tested whether these thresholds facilitate the differentiation between the B. melitensis field isolates and the Rev.1 vaccine strain (Table 2). The alignment procedure against a reference spectrum decreased the standard deviation considerably (Table 2), e.g., for "BfR" peaks within the ribosomal protein L24 (z = 1) decision mass window (11,197.5:11,235) from mean non−aligned = 11,206.9 with SD non−aligned = 2.9 for technical replicate spectra to mean aligned = 11,208.1 with SD aligned = 0.4 for sample spectra (Figures 3D-F Recently, a study has been published that also aimed to detect biomarkers enabling the discrimination of B. melitensis field isolates and the Rev.1 vaccine strain by MALDI-TOF MS (Christoforidou et al, 2020). For this purpose, 73 clinical and veterinary B. melitensis isolates from Greece were analyzed.…”

Section: Differentiation Of Brucella Melitensismentioning

confidence: 99%

“…from B. melitensis 16M (Figure 4B). Christoforidou et al (2020) probably did not detect these ribosomal biomarkers since their forward analysis tolerance parameter, which was not stated, may have excluded the detection of close peaks with the Mass-Up default settings or due to low mass intensities in m/z ranges above 10 kDa. In order to prove the universal character of our biomarkers and to exclude clonal and regional effects, B. melitensis strains from 17 different MLVA groups (Supplementary Table 6) were subjected to MALDI-TOF MS analysis and the spectra were searched for the presence of the protein biomarkers L24 and S12.…”

Section: Differentiation Of Brucella Melitensismentioning

confidence: 99%

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Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS

et al. 2021

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Brucella melitensis Rev.1 is a live attenuated vaccine strain that is widely used to control brucellosis in small ruminants. For successful surveillance and control programs, rapid identification and characterization of Brucella isolates and reliable differentiation of vaccinated and naturally infected animals are essential prerequisites. Although MALDI-TOF MS is increasingly applied in clinical microbiology laboratories for the diagnosis of brucellosis, species or even strain differentiation by this method remains a challenge. To detect biomarkers, which enable to distinguish the B. melitensis Rev.1 vaccine strain from B. melitensis field isolates, we initially searched for unique marker proteins by in silico comparison of the B. melitensis Rev.1 and 16M proteomes. We found 113 protein sequences of B. melitensis 16M that revealed a homologous sequence in the B. melitensis Rev.1 annotation and 17 of these sequences yielded potential biomarker pairs. MALDI-TOF MS spectra of 18 B. melitensis Rev.1 vaccine and 183 Israeli B. melitensis field isolates were subsequently analyzed to validate the identified marker candidates. This approach detected two genus-wide unique biomarkers with properties most similar to the ribosomal proteins L24 and S12. These two proteins clearly discriminated B. melitensis Rev.1 from the closely related B. melitensis 16M and the Israeli B. melitensis field isolates. In addition, we verified their discriminatory power using a set of B. melitensis strains from various origins and of different MLVA types. Based on our results, we propose MALDI-TOF MS profiling as a rapid, cost-effective alternative to the traditional, time-consuming approach to differentiate certain B. melitensis isolates on strain level.

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Section: Differentiation Of Brucella Melitensismentioning

confidence: 66%

Section: Differentiation Of Brucella Melitensismentioning

confidence: 66%

Section: Differentiation Of Brucella Melitensismentioning

confidence: 91%

Section: Differentiation Of Brucella Melitensismentioning

confidence: 99%

Section: Differentiation Of Brucella Melitensismentioning

confidence: 99%

See 3 more Smart Citations

Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS

et al. 2021

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Molecular characterization and antimicrobial susceptibility testing of clinical and non-clinical Brucella melitensis and Brucella abortus isolates from Egypt

et al. 2021

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A Combination of MALDI-TOF MS Proteomics and Species-Unique Biomarkers’ Discovery for Rapid Screening of Brucellosis

Hamidi

Nejad

Es‐haghi

et al. 2022

J. Am. Soc. Mass Spectrom.

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Brucellosis is considered to be a zoonotic infection with a predominant incidence in most parts of Iran that may even simply involve diagnostic laboratory personnel. In the present study, we apply matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS) for rapid and reliable discrimination of Brucella abortus and Brucella melitensis, based on proteomic mass patterns from chemically treated whole-cell analyses. Biomarkers of the low molecular weight proteome in the MALDI-TOF MS spectra were assigned to conserved ribosomal and structural protein families that were found in genome assemblies of B. abortus and B. melitensis in the NCBI database. Significant protein mass signals successfully mapped to ribosomal proteins and structural proteins, such as integration host factor subunit alpha, cold-shock proteins, HU family DNA-binding protein, ATP synthase subunit C, and GNAT family N-acetyltransferase, with specific biomarker peaks that have been identified for each virulent and vaccine strain. Web-accessible bioinformatics algorithms, with a robust data analysis workflow, followed by ribosomal and structural protein mapping, significantly enhanced the reliable assignment of key proteins and accurate identification of Brucella species. Furthermore, clinical samples were analyzed to confirm the most dominant protein biomarker candidates and their relevance for the identifications of B. melitensis and B. abortus. With proper optimization, we envision that the presented MALDI-TOF MS proteomics analyses, coupled with special usage of bioinformatics, could be used as a cost-efficient strategy for the diagnostics of brucellosis and introduce a reliable identification protocol for species of dangerous bacteria.

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Identification of Brucella spp. isolates and discrimination from the vaccine strain Rev.1 by MALDI-TOF mass spectrometry

Cited by 7 publications

References 21 publications

Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS

Protein Biomarker Identification for the Discrimination of Brucella melitensis Field Isolates From the Brucella melitensis Rev.1 Vaccine Strain by MALDI-TOF MS

Molecular characterization and antimicrobial susceptibility testing of clinical and non-clinical Brucella melitensis and Brucella abortus isolates from Egypt

A Combination of MALDI-TOF MS Proteomics and Species-Unique Biomarkers’ Discovery for Rapid Screening of Brucellosis

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