An alternative to DNA extraction for the diagnosis of Pneumocystis carinii pneumonia by polymerase chain reaction using a new oligonucleotide probe.
We have used new specific primers and probe in a polymerase chain reaction (PCR) followed by Southern blot assays to detect Pneumocystis carinii in human bronchoalveolar lavage samples obtained from HIV-infected patients with pulmonary symptoms. To facilitate the procedure we developed a filtration technique without DNA extraction yielding a high sensitivity (18/18 positive results). The high specificity of the technique was shown by testing immunosuppressed patients without P. carinii pneumonia.
Borensztein
L
L
INSERM U.13, Hôpital Claude Bernard, Paris, France.
Hatin
I
I
Simonpoli
A M
AM
Ugarte
E
E
Girard
P M
PM
Jaureguiberry
G
G
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D001992:Bronchoalveolar Lavage Fluid; D004271:DNA, Fungal / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics; D015658:HIV Infections / Q000150:complications; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D011010:Pneumocystis / Q000235:genetics / Q000302:isolation & purification; D011020:Pneumonia, Pneumocystis / Q000150:complications / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D051381:Rats; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013698:Templates, Genetic
D004271:DNA, Fungal; D004275:DNA, Ribosomal; D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S