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Rapid Diagnosis and Simultaneous Identification of Tuberculous and Bacterial Meningitis by a Newly Developed Duplex Polymerase Chain Reaction

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Abstract

The present study describes the development and evaluation of a duplex polymerase chain reaction (D-PCR) for diagnosis and simultaneous identification of tuberculous meningitis (TBM) and bacterial meningitis (BM) in a single reaction. A D-PCR with primers amplifying portions of the Mycobacterium tuberculosis IS6110 and the eubacteria 16SrDNA sequence in a same reaction mix was developed and tested on DNA extracted from 150 clinical CSF samples from different categories (TBM = 39, BM = 26, control infectious and non-infectious category = 85). The results indicate a clear differentiation between bands for eubacteria and M. tuberculosis with an analytical sensitivity of 103 cfu/ml for eubacteria and 102 cfu/ml for M. tuberculosis. When evaluated in clinical samples, D-PCR overall diagnosed 100 % confirmed TBM and 100 % confirmed BM cases with overall specificity of 96.5 %. D-PCR can be an effective tool for diagnosis and simultaneous identification of TBM or BM in a single PCR reaction. It saves time, cost, labour and sample amount and help in administration of appropriate antimicrobial therapy. The proposed diagnostic assay would be helpful in correct and rapid management of TBM and BM patients.

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Acknowledgments

The authors acknowledge Central India Institute of Medical Sciences, Nagpur, Maharashtra, India for providing laboratory working space and clinical data of cases.

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There is no conflict of interest to declare.

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Authors and Affiliations

  1. Biochemistry Research Laboratory, Central India Institute of Medical Sciences, 88/2, Bajaj Nagar, Nagpur, 440010, Maharashtra, India

    Khushboo J. Nagdev, Sharda P. Bhagchandani, Shradha S. Bhullar, Rajkumar C. Kapgate, Rajpal S. Kashyap, Nitin H. Chandak, Hatim F. Daginawala & Girdhar M. Taori

  2. Environmental Genomics Unit, National Environmental Engineering Research Institute, Nehru Marg, Nagpur, 440020, India

    Hemant J. Purohit

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  1. Khushboo J. Nagdev
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  2. Sharda P. Bhagchandani
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  3. Shradha S. Bhullar
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  4. Rajkumar C. Kapgate
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  5. Rajpal S. Kashyap
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  6. Nitin H. Chandak
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  7. Hatim F. Daginawala
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  8. Hemant J. Purohit
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  9. Girdhar M. Taori
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Corresponding author

Correspondence to Rajpal S. Kashyap.

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Nagdev, K.J., Bhagchandani, S.P., Bhullar, S.S. et al. Rapid Diagnosis and Simultaneous Identification of Tuberculous and Bacterial Meningitis by a Newly Developed Duplex Polymerase Chain Reaction. Indian J Microbiol 55, 213–218 (2015). https://doi.org/10.1007/s12088-015-0517-9

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  • DOI: https://doi.org/10.1007/s12088-015-0517-9

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