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Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods

Received: 30 April 2022     Accepted: 18 May 2022     Published: 9 June 2022
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Abstract

Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found < 1.7 ± 0, in milk 1.2 ± 0.2, in eggs < 1.2 ± 0.2, and in feed pellet < 1.6 ± 0.2 CFU/test portion. The accuracy, sensitivity, and specificity of the method were found 98%, 96%, and 100% respectively. No significant effects (p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin.

Published in International Journal of Microbiology and Biotechnology (Volume 7, Issue 2)
DOI 10.11648/j.ijmb.20220702.16
Page(s) 84-92
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2022. Published by Science Publishing Group

Keywords

Salmonella, Identification Method, Verification, MALDI TOF MS

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  • APA Style

    Md. Al-Amin, Md. Mizanur Rahman, Marufa Aktar, Md. Mostofa Kamal. (2022). Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. International Journal of Microbiology and Biotechnology, 7(2), 84-92. https://doi.org/10.11648/j.ijmb.20220702.16

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    ACS Style

    Md. Al-Amin; Md. Mizanur Rahman; Marufa Aktar; Md. Mostofa Kamal. Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. Int. J. Microbiol. Biotechnol. 2022, 7(2), 84-92. doi: 10.11648/j.ijmb.20220702.16

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    AMA Style

    Md. Al-Amin, Md. Mizanur Rahman, Marufa Aktar, Md. Mostofa Kamal. Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods. Int J Microbiol Biotechnol. 2022;7(2):84-92. doi: 10.11648/j.ijmb.20220702.16

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  • @article{10.11648/j.ijmb.20220702.16,
      author = {Md. Al-Amin and Md. Mizanur Rahman and Marufa Aktar and Md. Mostofa Kamal},
      title = {Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {7},
      number = {2},
      pages = {84-92},
      doi = {10.11648/j.ijmb.20220702.16},
      url = {https://doi.org/10.11648/j.ijmb.20220702.16},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20220702.16},
      abstract = {Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin.},
     year = {2022}
    }
    

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  • TY  - JOUR
    T1  - Development and Verification of a MALDI-TOF MS-Based Method for Rapid and Confirmatory Identification of Salmonella in Feed and Foods
    AU  - Md. Al-Amin
    AU  - Md. Mizanur Rahman
    AU  - Marufa Aktar
    AU  - Md. Mostofa Kamal
    Y1  - 2022/06/09
    PY  - 2022
    N1  - https://doi.org/10.11648/j.ijmb.20220702.16
    DO  - 10.11648/j.ijmb.20220702.16
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 84
    EP  - 92
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20220702.16
    AB  - Salmonella infection is one of the major global public health hazards and remains as an economic burden to both developed and developing countries through the costs associated with trade banning, as well as surveillance, prevention, and treatment of the disease. Confirmatory and rapid identification of Salmonella in feed and foods of animal origin is crucial for mitigating the associated burdens. The conventional methods for isolation and identification of Salmonella species are based on culture and biochemical tests and are very time-consuming requiring 10-11 days. Hence, these drawbacks of the conventional methods warrant a rapid method for confirmatory identification of Salmonella in feed and foods of animal origin. Therefore, a method based on matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) was developed and verified for its reliable use in the laboratory. For verification, Salmonella typhimurium (ATCC 14028) was used as the target bacteria. E. coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) were used as non-target bacteria. Salmonella typhimurium (ATCC 14028) produced black colonies with metallic sheen on BS agars, black colonies on XLD agars, and black-centered blue colonies on HE agars. MALDI TOF MS was found very efficient for confirmatory identification of Salmonella bacteria at the genus level without performing biochemical tests. Confirmatory identification of Salmonella could be made within four to five days after the commencement of the test. The expected limit of detection (eLOD50) of Salmonella in chicken and beef was found p = 0.400) of sample matrices were found on test results carried out by the developed method. Likewise, in terms of ruggedness, no significant effects of analyst change (p = 0.787), incubator change (p = 0.787), and day change (p = 0.242) were found in the test results. The method was found robust and could be used in the laboratory for rapid and confirmatory identification of Salmonella in feeds, and foods of animal origin.
    VL  - 7
    IS  - 2
    ER  - 

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Author Information
  • Department of Livestock Services, Quality Control Laboratory, Savar, Bangladesh

  • Department of Livestock Services, Quality Control Laboratory, Savar, Bangladesh

  • Department of Livestock Services, Quality Control Laboratory, Savar, Bangladesh

  • Department of Livestock Services, Quality Control Laboratory, Savar, Bangladesh

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