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M9480872.TXT
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1994-09-05
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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).