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Raman spectroscopy reveals distinct differences between two closely related bacterial strains, Mycobacterium indicus pranii and Mycobacterium intracellulare

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Abstract

A common technique used to differentiate bacterial species and to determine evolutionary relationships is sequencing their 16S ribosomal RNA genes. However, this method fails when organisms exhibit high similarity in these sequences. Two such strains that have identical 16S rRNA sequences are Mycobacterium indicus pranii (MIP) and Mycobacterium intracellulare. MIP is of significance as it is used as an adjuvant for protection against tuberculosis and leprosy; in addition, it shows potent anti-cancer activity. On the other hand, M. intracellulare is an opportunistic pathogen and causes severe respiratory infections in AIDS patients. It is important to differentiate these two bacterial species as they co-exist in immuno-compromised individuals. To unambiguously distinguish these two closely related bacterial strains, we employed Raman and resonance Raman spectroscopy in conjunction with multivariate statistical tools. Phenotypic profiling for these bacterial species was performed in a kinetic manner. Differences were observed in the mycolic acid profile and carotenoid pigments to show that MIP is biochemically distinct from M. intracellulare. Resonance Raman studies confirmed that carotenoids were produced by both MIP as well as M. intracellulare, though the latter produced higher amounts. Overall, this study demonstrates the potential of Raman spectroscopy in differentiating two closely related mycobacterial strains.

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Acknowledgements

We are grateful to the infrastructure provided by the AFM facility, Centre for BioSystems Science and Engineering, and Ms. Monisha Vasist for her expertise in AFM imaging. This study was aided by the infrastructural support provided by the DBT-IISc partnership program, DST-FIST and UGC Centre for Advanced Study to the Department of Biochemistry, IISc. SU acknowledges the J.C Bose Fellowship from DST. AS is a senior fellow of Wellcome Trust-DBT India Alliance and acknowledges funding by a DBT grant (BT/PR13522/COE/34/27/2015) for this work. The support of all the members of the laboratories of DpN and SU is highly appreciated.

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Authors and Affiliations

  1. Centre for BioSystems Science and Engineering, Indian Institute of Science, Bangalore, 560012, India

    Taru Verma, Siva Umapathy & Dipankar Nandi

  2. Centre for Infectious Disease Research, Indian Institute of Science, Bangalore, 560012, India

    Santosh Podder, Mansi Mehta, Amit Singh & Dipankar Nandi

  3. Department of Biochemistry, Indian Institute of Science, Bangalore, 560012, India

    Santosh Podder & Dipankar Nandi

  4. All India Institute of Medical Sciences, Bhopal, 462020, India

    Sarman Singh

  5. Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore, 560012, India

    Amit Singh

  6. Department of Inorganic and Physical Chemistry, Indian Institute of Science, Bangalore, 560012, India

    Siva Umapathy

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  1. Taru Verma
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Verma, T., Podder, S., Mehta, M. et al. Raman spectroscopy reveals distinct differences between two closely related bacterial strains, Mycobacterium indicus pranii and Mycobacterium intracellulare. Anal Bioanal Chem 411, 7997–8009 (2019). https://doi.org/10.1007/s00216-019-02197-z

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