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Journal of Clinical Microbiology, April 2001, p. 1638-1643, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1638-1643.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

PCR-Hybridization Assay for Mycobacterium avium Complex: Optimization of Detection in Peripheral Blood from Humans

Giulio Ferrario,1 Andrea Gori,1,* Agostino Rossi,2 Lidia Catozzi,1 Chiara Molteni,1 Giulia Marchetti,1 Alessandra Bandera,1 Maria Cristina Rossi,1 Anna Degli Esposti,1 and Fabio Franzetti1

Institute of Infectious Diseases and Tropical Medicine, "Luigi Sacco" Hospital, University of Milan, Milan,1 and Laboratory of Microbiology, "Circolo-Macchi Foundation" Hospital, University of Insubria, Varese,2 Italy

Received 21 August 2000/Returned for modification 19 September 2000/Accepted 23 January 2001

We evaluated the sensitivity of a DNA amplification test for the detection of Mycobacterium avium in blood samples using different blood components and different DNA extraction methods. M. avium-inoculated blood samples were processed to obtain separate blood components: peripheral blood mononuclear cells (PBMCs), polymorphonuclear cells (PMNCs), and whole-blood sodium dodecyl sulfate (SDS)-lysate pellets. The sensitivity for the detection of the lowest mycobacterial load (1 CFU/ml) was significantly greater (P < 0.01) with DNA extracted from SDS-lysate pellets than with DNA extracted from PBMCs or PMNCs. Subsequently, DNA extraction methods based on guanidine NaOH, and proteinase were compared. The sensitivity of the guanidine-based method was significantly greater (P < 0.01) than those of the others.

* Corresponding author. Mailing address: Institute of Infectious Diseases and Tropical Medicine, "Luigi Sacco" Hospital, University of Milan, Via G. B. Grassi, 74, 20157 Milan, Italy. Phone: 39 02 39042676. Fax: 39 02 3560805. E-mail: andrea.gori{at}unimi.it.

Journal of Clinical Microbiology, April 2001, p. 1638-1643, Vol. 39, No. 4
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.4.1638-1643.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Huang, T.-S., Lee, S. S.-J., Chen, Y.-S., Tu, H.-Z., Huang, W.-K., Liu, Y.-C. (2005). Discordant molecular characterization results in a Mycobacterium avium complex strain isolated from an AIDS patient. J Med Microbiol 54: 681-683 [Abstract] [Full Text]  


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