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Efficient method for mycobacterial DNA extraction in blood cultures aids rapid PCR identification of Mycobacterium tuberculosis and Mycobacterium avium

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Abstract

The study presented here evaluated the utility of several methods of extracting mycobacterial nucleic acids from positive blood culture samples and examined the effect of each method on the performance of an in-house PCR used directly in the peripheral blood of 80 patients with AIDS to identify Mycobacterium spp. The modified Boom method for extracting DNA from blood cultures proved to be the most efficient, with subsequent PCR analysis yielding 100% positivity (7 samples positive for M. avium and 5 for M. tuberculosis). Only three of 12 patients with a positive blood culture had a PCR result positive for M. avium in peripheral blood. The identification of mycobacteria by PCR in blood culture took about 3 days, reducing the time to diagnosis by several weeks. These results demonstrate that PCR is a sensitive and quick method for identifying mycobacteria, especially when a good DNA extraction method is applied.

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Acknowledgments

This study was approved by the ethics committee of the Hospital de Clínicas (Federal University of Paraná, Curitiba, Brazil). We would like to express our gratitude to LACEN-Instituto de Saúde do Paraná for supporting this investigation.

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Authors and Affiliations

  1. LACEN-PR (Laboratório Central do Estado), Ubaldino do Amaral, 545 Alto da XV, Curitiba-PR, CEP-80060-190, Brazil

    S. M. Nakatani, M. Burger & S. R. Brockelt

  2. Department of Infectology, Federal University of Paraná, R Padre Camargo 280, Curitiba-PR, 80069-900, Brazil

    M. C. Assef

  3. Laboratory of Bacteriology, University Hospital of the Federal University of Paraná, R Padre Camargo 280, Curitiba-PR, 80069-900, Brazil

    L. L. Cogo

  4. Department of Clinical Pathology, Federal University of Paraná, R Padre Camargo 280, Curitiba-PR, 80069-900, Brazil

    I. J. T. Messias-Reason

Authors
  1. S. M. Nakatani
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  2. M. Burger
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  3. M. C. Assef
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  4. S. R. Brockelt
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  5. L. L. Cogo
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  6. I. J. T. Messias-Reason
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Corresponding author

Correspondence to I. J. T. Messias-Reason.

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Nakatani, S.M., Burger, M., Assef, M.C. et al. Efficient method for mycobacterial DNA extraction in blood cultures aids rapid PCR identification of Mycobacterium tuberculosis and Mycobacterium avium. Eur J Clin Microbiol Infect Dis 23, 851–854 (2004). https://doi.org/10.1007/s10096-004-1236-z

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  • DOI: https://doi.org/10.1007/s10096-004-1236-z

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