Detection of human Pneumocystis carinii by the polymerase chain reaction.
Oligonucleotide primers were prepared from a clone (B12) which has been shown to be a repetitive sequence in the rat P. carinii genome. Polymerase chain reaction was employed to amplify both rat and human P. carinii DNA. The detection limit of the assay was approximately 600 ng of total nucleic acid. Amplification products from both the rat and human isolates (ca. 780 bp) were characterized by denaturing gradient gel electrophoresis after digestion with Sau3A. No amplification products were obtained when DNA from the following potential pulmonary pathogens were used in identical reactions: Aspergillus fumigatus, Cryptococcus neoformans, Candida albicans, Mycobacterium avium-intracellulare and cytomegalovirus. In a blind study using the B12 primers, P. carinii DNA was successfully amplified in clinical samples which were positive by direct immunofluorescence assay (IFA) as well as in some specimens not identified by direct IFA.
Becker-Hapak
M
M
Department of Microbiology and Immunology, University of Oklahoma Health Science Center, Oklahoma City 73190.
Liberator
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P
Graves
D
D
D000818:Animals; D001483:Base Sequence; D004271:DNA, Fungal / Q000302:isolation & purification; D006801:Humans; D008168:Lung / Q000382:microbiology; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D051381:Rats; D011919:Rats, Inbred Strains
D004271:DNA, Fungal