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Simple Identification of Mycobacterial Species by Sequence-Specific Multiple Polymerase Chain Reactions

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Abstract

Several species of mycobacteria cause infections in humans. Species identification of clinical isolates of mycobacteria is very important for the decision of treatment and in choosing the appropriate treatment regimen. We have developed a multiplex PCR method that can identify practically all known species of mycobacteria, by determination of single-nucleotide differences at a total of 13 different polymorphic regions in the genes of rRNA and hsp65, in four PCR mixes. To achieve this goal, single-nucleotide differences in these polymorphic regions were used to divide mycobacterial species into two groups, than four, eight, etc., in an algorithmic manner. It was sufficient to reach single species level by evaluating 13 polymorphic regions. Evaluation of the multiplex PCR patterns by observable real-time electrophoresis (ORTE) simplified species identification. This new method may enable easy, rapid, and cost-effective identification of all species of mycobacteria.

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Acknowledgements

This work was supported by TUBİTAK 107S014 (SBAG-3541) Identification of Mycobacterial Species by Sequence-Specific Polymerase Chain Reaction; for financial aid, I wish to thank TÜBİTAK.

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Authors and Affiliations

  1. Department of Medical Microbiology, Faculty of Medicine, Acibadem Mehmet Ali Aydınlar University, Istanbul, Turkey

    Nihan Unubol, Sinem Oktem Okullu & Tanil Kocagoz

  2. Department of Medical Biotechnology, Institute of Health Sciences, Acibadem Mehmet Ali Aydınlar University, Istanbul, Turkey

    Nihan Unubol, Sinem Oktem Okullu & Tanil Kocagoz

  3. Faculty of Sciences, Department of Biology, Ege University, Izmir, Turkey

    Inci Tuney Kizilkaya

  4. Aziz Sancar Institute of Experimental Medicine, Istanbul University, Istanbul, Turkey

    Kaya Koksalan

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  1. Nihan Unubol
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  2. Inci Tuney Kizilkaya
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  3. Sinem Oktem Okullu
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  4. Kaya Koksalan
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  5. Tanil Kocagoz
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Correspondence to Tanil Kocagoz.

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Unubol, N., Kizilkaya, I.T., Okullu, S.O. et al. Simple Identification of Mycobacterial Species by Sequence-Specific Multiple Polymerase Chain Reactions. Curr Microbiol 76, 791–798 (2019). https://doi.org/10.1007/s00284-019-01661-4

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  • DOI: https://doi.org/10.1007/s00284-019-01661-4

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