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PCR-RFLP analysis of fliC, fimH and 16S rRNA genes in Salmonella Typhimurium isolates of varied origin

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Abstract

Restricted fragment length polymorphism (RFLP) was used in analyses on the typing and heterogeneity, typeability and polymorphism of the 16S rRNA, fliC and fimH genes in Salmonella Typhimurium isolates of varied origin. The digestion of PCR products with restriction enzymes EcoRV, ClaI, HaeIII and ScaI (fliC genes), HincII, ClaI, EcoRV and MluI (fimH genes) and EcoRI, SmaI and HaeIII (16S rRNA genes) generated two to four bands of ranging in size from 100 to 1,104 bp. Of all the restriction profiles obtained, only the ClaI profile for fimH could be used to classify Salmonella Typhimurium isolates into different groups. According to this profile, pattern A with uncut fimH was observed in eight isolates (36.36 %) and pattern B with 755- and 253-bp bands was observed in 14 isolates (63.63 %). No pattern was allotted for a special region or source. These results demonstrate that PCR-RFLP based on these genes showed good typeability but low discriminatory power. Moreover, the highly conserved nature of fliC, fimH and 16S rRNA illustrated in our study suggests the importance of these genes as immunization and diagnostic factors in Salmonella Typhimurium. Simultaneously, our results also illustrate the potential of ClaI-based fimH analysis as a marker for the sub-serotype level differentiation of Salmonella Typhimurium isolates.

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Acknowledgements

The authors are grateful to the Director and Joint Director (Academic and Research) of the Institute for providing the necessary research facilities to carry out the present investigation. TGS thanks the Indian Council of Agricultural Research (ICAR) for providing financial support in the form of a Junior Research Fellowship.

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Authors and Affiliations

  1. Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243122, India

    Thangalazhy Gopakumar Sumithra, Vinod Kumar Chaturvedi, Praveen Kumar Gupta, Ajay Rai, Sunita Chougule, Bincy Joseph, Susan Cherian & Chabi Lal Patel

  2. Division of Biological Products, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, 243122, India

    Vinod Kumar Chaturvedi

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  1. Thangalazhy Gopakumar Sumithra
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  2. Vinod Kumar Chaturvedi
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  5. Sunita Chougule
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Correspondence to Vinod Kumar Chaturvedi.

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Sumithra, T.G., Chaturvedi, V.K., Gupta, P.K. et al. PCR-RFLP analysis of fliC, fimH and 16S rRNA genes in Salmonella Typhimurium isolates of varied origin. Ann Microbiol 64, 177–183 (2014). https://doi.org/10.1007/s13213-013-0650-9

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  • DOI: https://doi.org/10.1007/s13213-013-0650-9

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