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.

PCR-hybridization assay for Mycobacterium avium complex: optimization of detection in peripheral blood from humans / G. Ferrario, A. Gori, A. Rossi, L. Catozzi, C. Molteni, G. Marchetti, A. Bandera, M.C. Rossi, A. Degli Esposti, F. Franzetti. - In: JOURNAL OF CLINICAL MICROBIOLOGY. - ISSN 0095-1137. - 39:4(2001 Apr), pp. 1638-1643.

PCR-hybridization assay for Mycobacterium avium complex: optimization of detection in peripheral blood from humans

G. Ferrario;A. Gori;G. Marchetti;A. Bandera;
2001-04

Abstract

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.
human-immunodeficiency-virus; polymerase chain-reaction; clinical specimens; identification; tuberculosis; bacteremia; infection; disease; AIDS; diagnosis
Settore MED/17 - Malattie Infettive
JOURNAL OF CLINICAL MICROBIOLOGY
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/22637
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