PCR-hybridization assay for Mycobacterium avium complex: optimization of detection in peripheral blood from humans.
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.
Institute of Infectious Diseases and Tropical Medicine, Luigi Sacco Hospital, University of Milan, Milan, Varese, Italy.
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Ferrario
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Gori
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Rossi
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L
Catozzi
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Molteni
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Marchetti
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Bandera
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Rossi
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Esposti
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Franzetti
D016470:Bacteremia / Q000382:microbiology; D001769:Blood / Q000382:microbiology*; D004269:DNA, Bacterial / Q000097:blood* / Q000302:isolation & purification; D006801:Humans; D015269:Mycobacterium avium Complex / Q000235:genetics / Q000302:isolation & purification*; D015270:Mycobacterium avium-intracellulare Infection / Q000382:microbiology*; D009693:Nucleic Acid Hybridization / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial