Detection and identification of Mycobacterium avium in the blood of AIDS patients by the polymerase chain reaction.
One hundred fifty-three blood samples from patients positive for the human immunodeficiency virus (HIV) were analyzed by polymerase chain reaction (PCR) to detect the presence of Mycobacterium avium. Samples were collected from patients who also had blood cultures performed by a radiometric method. Blood samples were centrifuged on a Ficoll-Hypaque gradient to purify peripheral blood mononuclear cells. The purified cells were washed and incubated with a resin, boiled to release mycobacterial DNA, and then amplified. Polymerase chain reaction products were detected by a nonisotopic method. A 123 base-pair (bp) insertion sequence, namely IS6110, from Mycobacterium tuberculosis complex was also included in the reaction as an internal control of Taq polymerase activity to exclude the presence of enzyme inhibitors. This IS6110 fragment can be distinguished from the 383 bp target product on ethidium bromide-stained agarose gel and may also be used in a colorimetric assay. Such results were compared with the results of culture and indicated that the assay is as sensitive as bacteriological methods, though faster.
De Francesco
M A
MA
Institute of Microbiology, University of Brescia, Italy.
Colombrita
D
D
Pinsi
G
G
Gargiulo
F
F
Caligaris
S
S
Bertelli
D
D
Martinelli
F
F
Gao
J
J
Turano
A
A
D000163:Acquired Immunodeficiency Syndrome / Q000150:complications*; D003429:Cross Reactions; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D009162:Mycobacterium avium / Q000302:isolation & purification*; D015269:Mycobacterium avium Complex / Q000235:genetics; D015270:Mycobacterium avium-intracellulare Infection / Q000097:blood / Q000150:complications / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*