The ability of MALDI-TOF MS to identify Salmonella isolated from food
Main Article Content
Abstract
Salmonella is among the leading foodborne pathogens in the European Union, with Salmonella Enteritidis and Salmonella Typhimurium being the most frequently reported serovars. Traditional methods for Salmonella identification and serotyping, such as ISO standards, are time-consuming and labor-intensive. MALDI-TOF MS has emerged as a rapid and reliable tool for bacterial identification. This study evaluated the ability of MALDI-TOF MS in identifying Salmonella isolates collected from various stages of chicken and pork meat production. Eight pathogenic isolates (four S. Enteritidis and four S. Typhimurium) were analyzed using both direct colony transfer and protein extraction methods. Results showed that the direct transfer method yielded low-confidence or no identification for four isolates. However, all isolates were successfully identified with high-confidence scores (> 2.00) after protein extraction. Despite the good scores, all isolates were identified only at the genus level (Salmonella sp.), consistent with current limitations of the MALDI Biotyper database. Nevertheless, best database matches after protein extraction indicated potential for more detailed classification. This study confirms that MALDI-TOF MS, particularly when combined with protein extraction, is a valuable method for rapid screening and identification of Salmonella sp. in food production chain. However, its inability to perform accurate serotyping highlights the need for improved databases and computational approaches. Future integration of machine learning and expanded reference spectra may enhance the serotype-level resolution of MALDI-TOF MS.
Downloads
Article Details

This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors retain copyright of the published papers and grant to the publisher the right to publish the article, to be cited as its original publisher in case of reuse, and to distribute it in all forms and media. Articles will be distributed under the Creative Commons Attribution International License (CC BY 4.0).
References
Alatoom A. A., Cunningham S. A., Ihde S. M., Mandrekar J., Patel, R. 2011. Comparison of direct colony method versus extraction method for identification of Gram-positive cocci by use of Bruker Biotyper matrix-assisted laser desorption ionization-time of flight mass spectrometry. Journal of Clinical Microbiology, 49 (8):2868-2873. https://doi.org/10.1128/jcm.00506-11
Al-Hindi R. R., Alharbi M. G., Alotibi I. A., Azhari S. A., Ahmad A., Alseghayer M. S.,Teklemariam A. D., Almaneea, A. M. 2023. MALDI-TOF MS-based identification and antibiotics profiling of Salmonella species isolated from retail chilled chicken in Saudi Arabia. Journal of King Saud University-Science, 35(5), 102684. https://doi.org/10.1016/j.jksus.2023.102684
Anderson N. W., Buchan B. W., Riebe K. M., Parsons L. N., Gnacinski S., Ledeboer N. A. 2012. Effects of solid-medium type on routine identification of bacterial isolates by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry. Journal of Clinical Microbiology, 50(3):1008-1013. https://doi.org/10.1128/jcm.05209-11.
Bastin B., Bird P., Benzinger M. J., Crowley E., Agin J., Goins D., Sohier D., Timke M., Shi G., Kostrzewa M. 2019. Confirmation and Identification of Salmonella spp., Cronobacter spp., and Other Gram-Negative Organisms by the Bruker MALDI Biotyper Method: Collaborative Study Method Extension to Include Campylobacter Species, Revised First Action 2017.09. Journal of AOAC International, 102(5):1595-1616. https://doi.org/10.1093/jaoac/102.5.1595
Böhme K., Antelo S. C., Fernández-No I. C., Quintela-Baluja M., Barros-Velázquez J., Cañas, B. Calo-Mata, P. 2016. Detection of foodborne pathogens using MALDI-TOF mass spectrometry. In Antimicrobial Food Packaging, Academic Press: Barros-Velázquez, J. https://doi.org/10.1016/B978-0-12-800723-5.00015-2
Dieckmann R., Malorny B. 2011. Rapid screening of epidemiologically important Salmonella enterica subsp. enterica serovars by whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry. Applied and Environmental Microbiology, 77(12):4136-4146. https://doi.org/10.1128/AEM.02418-10
Dieckmann R., Helmuth R., Erhard M., Malorny B. 2008. Rapid classification and identification of Salmonellae at the species and subspecies levels by whole-cell matrix-assisted laser desorption ionization-time of flight mass spectrometry. Applied and Environmental Microbiology, 74(24):7767-7778. https://doi.org/10.1128/AEM.01402-08
Elbehiry A., Marzouk E., Hamada M., Al-Dubaib M., Alyamani E., Moussa I. M., AlRowaidhan A., Hemeg H. A. 2017. Application of MALDI-TOF MS fingerprinting as a quick tool for identification and clustering of foodborne pathogens isolated from food products. New Microbiologica, 40(4):269-278.
European Food Safety Authority (EFSA), & European Centre for Disease Prevention and Control (ECDC). 2024. The European Union One Health 2023 Zoonoses Report. EFSA Journal, 22(12), e9106.
International Organization for Standardization (ISO). 2017. Microbiology of the food chain-horizontal method for the detection, enumeration and serotyping of Salmonella-Part 1: detection of Salmonella spp. ISO 6579-1:2017. Geneva, Switzerland.
Jadhav S. R., Shah R. M., Karpe A. V., Morrison P. D., Kouremenos K., Beale D. J., Palombo E. A. 2018. Detection of foodborne pathogens using proteomics and metabolomics-based approaches. Frontiers in Microbiology, 9, 3132. https://doi.org/10.3389/fmicb.2018.03132
Kang L., Li N., Li P., Zhou Y., Gao S., Gao H., Yin W., Wang J. 2017. MALDI-TOF mass spectrometry provides high accuracy in identification of Salmonella at species level but is limited to type or subtype Salmonella serovars. European Journal of Mass Spectrometry, 23(2):70-82. https://doi.org/10.1177/1469066717699216
Mangmee S., Reamtong O., Kalambaheti T., Roytrakul S., Sonthayanon P. 2020. MALDI-TOF mass spectrometry typing for predominant serovars of non-typhoidal Salmonella in a Thai broiler industry. Food Control, 113, 107188. https://doi.org/10.1016/j.foodcont.2020.107188
Mazzeo M. F., Sorrentino A., Gaita M., Cacace G., Di Stasio M., Facchiano A., Comi G., Malorni A., Siciliano R. A. 2006. Matrix-assisted laser desorption ionization-time of flight mass spectrometry for the discrimination of food-borne microorganisms. Applied and Environmental Microbiology, 72(2):1180-1189. https://doi.org/10.1128/AEM.72.2.1180-1189.2006
Mkangara M. 2023. Prevention and control of human Salmonella enterica infections: An implication in food safety. International Journal of Food Science, 2023(1), 8899596. https://doi.org/10.1155/2023/8899596
Pavlović M., Huber I., Konrad R., Busch U. 2013. Application of MALDI-TOF MS for the identification of food borne bacteria. The Open Microbiology Journal, 7:135-141. https://doi.org/10.2174/1874285801307010135
Persad A. K., Fahmy H. A., Anderson N., Cui J., Topalcengiz Z., Jeamsripong S., ... LeJeune J. T. 2022. Identification and subtyping of Salmonella isolates using matrix-assisted laser desorption–ionization time-of-flight mass spectrometry (MALDI-TOF). Microorganisms, 10(4):688. https://doi.org/10.3390/microorganisms10040688
Ren J., Xia J., Zhang M., Liu C., Xu Y., Wu J., ... Cao W. 2025. Automated identification of Salmonella serotype using MALDI-TOF mass spectrometry and machine learning techniques. Journal of Clinical Microbiology, e00037-25. https://doi.org/10.1128/jcm.00037-25
Ryzhov V., Fenselau C. 2001. Characterization of the protein subset desorbed by MALDI from whole bacterial cells. Analytical Chemistry, 73(4):746-750. https://doi.org/10.1021/ac0008791
Sparbier K., Weller U., Boogen C., Kostrzewa M. 2012. Rapid detection of Salmonella sp. by means of a combination of selective enrichment broth and MALDI-TOF MS. European Journal of Clinical Microbiology & Infectious Diseases, 31:767-773. https://doi.org/10.1007/s10096-011-1373-0
Vidaković Knežević S., Pelić M., Žekić M., Knežević S., Vranešević J., Ljubojević Pelić D., Savić S. 2024. The presence of Salmonella enterica and Listeria monocytogenes on poultry slaughter plant equipment after sanitation procedures. Archives of Veterinary Medicine, 17(2):115-125. https://doi.org/10.46784/e-avm.v17i2.402
Wenning M., Breitenwieser F., Konrad R., Huber I., Busch U., Scherer S. 2014. Identification and differentiation of food-related bacteria: a comparison of FTIR spectroscopy and MALDI-TOF mass spectrometry. Journal of Microbiological Methods, 103:44-52. https://doi.org/10.1016/j.mimet.2014.05.011
Wieser A., Schneider L., Jung J., Schubert S. 2012. MALDI-TOF MS in microbiological diagnostics – identification of microorganisms and beyond (mini review). Applied Microbiology and Biotechnology, 93:965-974. https://doi.org/10.1007/s00253-011-3783-4
Yan W., Qian J., Ge Y., Ye K., Zhou C., Zhang H. 2020. Principal component analysis of MALDI-TOF MS of whole-cell foodborne pathogenic bacteria. Analytical Biochemistry, 592, 113582. https://doi.org/10.1016/j.ab.2020.113582