Document 0872
 DOCN  M9480872
 TI    Polymerase chain reaction (PCR) detection of Mycobacterium
       avium-intracellulare (MAI) DNA from peripheral blood mononuclear cells
       (PBMCs) of HIV-infected patients.
 DT    9410
 AU    Kostman J; Mair M; Byrne B; Gekowski K; Baxter J; Cooper Hospital/UMC,
       Robert Wood Johnson Medical School, Camden,; N.J.
 SO    Abstr Gen Meet Am Soc Microbiol. 1994;94:188 (abstract no. U-91). Unique
       Identifier : AIDSLINE ASM94/94313073
 AB    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.
 DE    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  MEETING ABSTRACT

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).