Detection of Pneumocystis carinii in serum of AIDS patients with Pneumocystis pneumonia by the polymerase chain reaction.
Amplification of DNA by the polymerase chain reaction (PCR) offers a highly sensitive and specific method for detecting DNA sequences in biological samples. We applied this technology to develop an assay for the P. carinii dihydrofolate reductase (DHFR) gene. This assay was found to be sensitive enough to detect as little as 1 organism-'equivalent' of DHFR DNA. In rats with experimentally-induced P. carinii pneumonia, DHFR DNA amplification demonstrated the presence of pulmonary P. carinii 2 wk prior to the onset of histopathological changes. When rat serum was analyzed by PCR, serum P. carinii DNA was found in 5 of 14 experimental rats. Finally, P. carinii DNA was detected in the serum of 7 of 18 patients (39%) with AIDS and active P. carinii pneumonia. These results suggest that circulating serum P. carinii DNA can be detected frequently in the course of pulmonary infection and may represent a blood-borne phase of infection. The PCR detection of P. carinii DNA provides a useful tool to study the natural history of P. carinii infection and may offer a non-invasive diagnostic procedure in some patients with P. carinii pneumonia.
Schluger
N
N
New York Hospital-Cornell Medical Center, NY 10021.
Sepkowitz
K
K
Armstrong
D
D
Bernard
E
E
Rifkin
M
M
Cerami
A
A
Bucala
R
R
D000163:Acquired Immunodeficiency Syndrome / Q000097:blood / Q000150:complications*; D000818:Animals; D001483:Base Sequence; D005800:Genes, Fungal; D008168:Lung / Q000382:microbiology; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000201:enzymology / Q000302:isolation & purification*; D011020:Pneumonia, Pneumocystis / Q000097:blood / Q000150:complications / Q000382:microbiology*; D016133:Polymerase Chain Reaction; D051381:Rats; D013762:Tetrahydrofolate Dehydrogenase / Q000235:genetics
D013762:Tetrahydrofolate Dehydrogenase