PCR-Hybridization Assay for Mycobacterium avium Complex: Optimization of Detection in Peripheral Blood from Humans
- 1 April 2001
- journal article
- research article
- Published by American Society for Microbiology in Journal of Clinical Microbiology
- Vol. 39 (4) , 1638-1643
- https://doi.org/10.1128/jcm.39.4.1638-1643.2001
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.Keywords
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