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Evaluation of Genetic Content of the CRISPR Locus in Listeria monocytogenes Isolated From Clinical, Food, Seafood and Animal Samples in Iran

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Abstract

CRISPR arrays, which are organized to fight against non-self DNA elements, have shown sequence diversity that could be useful in evolution and typing studies. In this study, 55 samples of L. monocytogenes isolated from different sources were evaluated for CRISPR sequence polymorphism. The CRISPR loci were identified using CRISPR databases. A single PCR assay was designed to amplify the target CRISPRs using an appropriate universal primer. Sequencing results were analyzed using CRISPR databases and BLASTn, and the CRISPR locus was present in all the strains. Three hundred repeats including 55 terminal repeats were identified. Four types of consensuses direct repeat (DR) with different lengths and sequences were characterized. Sixty repeat variants were observed which possessed different polymorphisms. Two hundred and fifty spacers were identified from which 35 consensus sequences were determined, indicating the high polymorphism of the CRISPR spacers. The identified spacers showed similarities to listeria phage sequences, other bacterial phage sequences, plasmid sequences and bacterial sequences. In order to control the bacterial outbreaks, a robust and precise system of subtyping is required. High levels of polymorphism in the CRISPR loci in this study might be related to the origin and time of the samples’ isolation. However, it is essential to assess, on a case-by-case basis, the characteristics of any given CRISPR locus before its use as an epidemiological marker. In conclusion, the results of this study showed that the use of sequence content of CRISPR area could provide new and valuable information on the evolution and typing of the L. monocytogenes bacterium.

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Acknowledgements

This study supported by Iran University of Medical Sciences, Tehran, IR Iran.

Funding

This study supported by Iran University of Medical Sciences, Tehran, IR Iran.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Medicine, Iran University of Medical Sciences, Hemmat Highway, Tehran, Iran

    Elnaz Ohadi & Shabnam Razavi

  2. Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran

    Asaad Azarnezhad

  3. Department of Microbiology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran

    Lida Lotfollahi

  4. Department of Microbiology, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran

    Parisa Asadollahi, Vahab Hasan Kaviar & Behrooz Sadeghi Kalani

  5. Clinical Microbiology Research Center, Ilam University of Medical Sciences, Ilam, Iran

    Parisa Asadollahi, Vahab Hasan Kaviar & Behrooz Sadeghi Kalani

  6. Microbial Biotechnology Research Center, Iran University of Medical Sciences, Tehran, Iran

    Shabnam Razavi

Authors
  1. Elnaz Ohadi
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  2. Asaad Azarnezhad
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  3. Lida Lotfollahi
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  4. Parisa Asadollahi
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  5. Vahab Hasan Kaviar
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  6. Shabnam Razavi
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  7. Behrooz Sadeghi Kalani
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Contributions

BSK and SR conceived and designed research. EO and AA conducted experiments. PA and LL analyzed data. VH wrote the manuscript. BSK and SR made a critical revision and contributed in writing. BSK and EO writing and finalizing of the article draft. All authors read and approved the manuscript.

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Correspondence to Shabnam Razavi or Behrooz Sadeghi Kalani.

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Ohadi, E., Azarnezhad, A., Lotfollahi, L. et al. Evaluation of Genetic Content of the CRISPR Locus in Listeria monocytogenes Isolated From Clinical, Food, Seafood and Animal Samples in Iran. Curr Microbiol 80, 388 (2023). https://doi.org/10.1007/s00284-023-03508-5

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  • DOI: https://doi.org/10.1007/s00284-023-03508-5

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