Polymerase chain reaction (PCR) detection of Mycobacterium avium-intracellulare (MAI) DNA from peripheral blood mononuclear cells (PBMCs) of HIV-infected patients.

Important note: Information in this article was accurate in 1994. The state of the art may have changed since the publication date.

Polymerase chain reaction (PCR) detection of Mycobacterium avium-intracellulare (MAI) DNA from peripheral blood mononuclear cells (PBMCs) of HIV-infected patients.

Abstr Gen Meet Am Soc Microbiol. 1994;94:188 (abstract no. U-91). Unique Identifier : AIDSLINE ASM94/94313073
Kostman J; Mair M; Byrne B; Gekowski K; Baxter J; Cooper Hospital/UMC, Robert Wood Johnson Medical School, Camden,; N.J.

Abstract: Rapid detection and identification of MAI from the blood of HIV-infected patients is necessary for the prompt initiation of treatment. We performed DNA amplification with the polymerase chain reaction of mycobacterial DNA from frozen PBMCs collected from HIV-infected patients. Heparinized blood was collected from HIV-infected patients (CD4 counts < 90/mm3) and PMBCs were prepared using a standard Ficoll-Paque separation and stored at 70 degrees C until further analysis. DNA was isolated from cells after lysis with lysozyme, digestion with proteinase K, phenol-chloroform extraction, and ethanol precipitation. Two sets of oligonucleotide primers based on 16S ribosomal RNA sequences were used to amplify any mycobacterial sequences. One primer pair amplified DNA from any mycobacterial species; internal primers were then used to amplify and distinguish DNA from either M. avium (194 base pairs) or M. intracellulare (519 base pairs). All samples were analyzed by PCR in a blinded fashion. We detected mycobacterial DNA from 9 of 14 samples of stored PBMCs, representing 7 patients. Two patients who subsequently had positive blood cultures for MAI had mycobacterial DNA detected from PBMCs at 8 and 12 months prior to the time of the positive cultures. These patients had symptoms compatible with disseminated MAI at the time the mycobacterial DNA was detected. No patients with positive blood cultures were negative by PCR. All positive samples were determined to be M. avium. The prompt detection of mycobacterial DNA from PBMCs of HIV-infected patients with low CD4 counts offers a sensitive, cost-effective alternative to routine culture techniques. Detection of disseminated infection with MAI at an earlier stage may lead to improved therapeutic responses.

Keywords: AIDS-Related Opportunistic Infections/BLOOD/*MICROBIOLOGY Cells, Cultured DNA Primers DNA, Bacterial/*BLOOD/ISOLATION & PURIF Human Monocytes/*MICROBIOLOGY Mycobacterium avium Complex/GENETICS/*ISOLATION & PURIF Polymerase Chain Reaction/*METHODS Single-Blind Method Tuberculosis/BLOOD/COMPLICATIONS/*DIAGNOSIS ABSTRACT
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M94A0881

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