Laboratory diagnosis of Pneumocystis carinii infections by PCR directed to genes encoding for mitochondrial 5S and 28S ribosomal RNA.
PCR with 5S mitochondrial ribosomal RNA (5S) target is a sensitive and specific assay for the detection of Pneumocystis carinii in clinical specimens from the respiratory tract. We developed an oligonucleotide probe directed to a 200 bp amplicon generated by fungal-specific universal primers that anneals with sequences specific for P. carinii in the 28S ribosomal RNA gene (28S). Of 50 archived bronchoalveolar lavage 1(BAL) specimens, 46 of 50 samples (92% agreement) gave the same result (23 positive, 23 negative) by PCR directed to the 5S and 28S assays. Results of calcofluor white staining of BAL smears on slides indicated agreement with the molecular results in 43 of 46 (93.5%) assays. PCR detection of P. carinii by amplification of 28S ribosomal gene target by fungal-specific primers and an organism-specific probe provides an alternate genomic target for the laboratory diagnosis of this organism.
Sandhu
G S
GS
Department of Biochemistry and Molecular Biology, Mayo Clinic and Foundation, Rochester, MN 55905, USA.
Kline
B C
BC
Espy
M J
MJ
Stockman
L
L
Smith
T F
TF
Limper
A H
AH
D000368:Aged; D001557:Benzenesulfonates; D001992:Bronchoalveolar Lavage Fluid / Q000382:microbiology; D004272:DNA, Mitochondrial; D005260:Female; D020459:Genes, rRNA; D006801:Humans; D008297:Male; D008875:Middle Aged; D011010:Pneumocystis / Q000235:genetics / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012331:RNA, Fungal / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling
D001557:Benzenesulfonates; D004272:DNA, Mitochondrial; D012331:RNA, Fungal; D012339:RNA, Ribosomal, 28S; D012341:RNA, Ribosomal, 5S; C007061:C.I. Fluorescent Brightening Agent 28