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

  • S. M. Nakatani
  • M. Burger
  • M. C. Assef
  • S. R. Brockelt
  • L. L. Cogo
  • I. J. T. Messias-Reason
Concise Article

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.

Keywords

Tuberculosis Blood Culture Benzyl Alcohol Positive Blood Culture Guanidine Hydrochloride 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

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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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • S. M. Nakatani
    • 1
  • M. Burger
    • 1
  • M. C. Assef
    • 2
  • S. R. Brockelt
    • 1
  • L. L. Cogo
    • 3
  • I. J. T. Messias-Reason
    • 4
  1. 1.LACEN-PR (Laboratório Central do Estado)Ubaldino do AmaralCuritiba-PRBrazil
  2. 2.Department of InfectologyFederal University of ParanáCuritiba-PRBrazil
  3. 3.Laboratory of BacteriologyUniversity Hospital of the Federal University of ParanáCuritiba-PRBrazil
  4. 4.Department of Clinical PathologyFederal University of ParanáCuritiba-PRBrazil

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