Detection and identification ofMycobacterium avium in the blood of AIDS patients by the polymerase chain reaction

  • M. A. De Francesco
  • D. Colombrita
  • G. Pinsi
  • F. Gargiulo
  • S. Caligaris
  • D. Bertelli
  • F. Martinelli
  • J. Gao
  • A. Turano
Article

DOI: 10.1007/BF01709362

Cite this article as:
De Francesco, M.A., Colombrita, D., Pinsi, G. et al. Eur. J. Clin. Microbiol. Infect. Dis. (1996) 15: 551. doi:10.1007/BF01709362

Abstract

One hundred fifty-three blood samples from patients positive for the human immunodeficiency virus (HIV) were analyzed by polymerase chain reaction (PCR) to detect the presence ofMycobacterium avium. Samples were collected from patients who also had blood cultures performed by a radiometric method. Blood samples were centrifuged on a Ficoll-Hypaque gradient to purify peripheral blood mononuclear cells. The purified cells were washed and incubated with a resin, boiled to release mycobacterial DNA, and then amplified. Polymerase chain reaction products were detected by a nonisotopic method. A 123 base-pair (bp) insertion sequence, namely IS6110, fromMycobacterium tuberculosis complex was also included in the reaction as an internal control ofTaq polymerase activity to exclude the presence of enzyme inhibitors. This IS6110 fragment can be distinguished from the 383 bp target product on ethidium bromide-stained agarose gel and may also be used in a colorimetric assay. Such results were compared with the results of culture and indicated that the assay is as sensitive as bacteriological methods, though faster.

Copyright information

© MMV Medizin Verlag GmbH 1996

Authors and Affiliations

  • M. A. De Francesco
    • 1
  • D. Colombrita
    • 1
  • G. Pinsi
    • 1
  • F. Gargiulo
    • 1
  • S. Caligaris
    • 2
  • D. Bertelli
    • 2
  • F. Martinelli
    • 1
  • J. Gao
    • 1
  • A. Turano
    • 1
  1. 1.Institute of MicrobiologyUniversity of BresciaBresciaItaly
  2. 2.II and I Division of Infectious DiseasesSpedali CiviliItaly

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