Comparative study of antipneumocystis agents in rats by using a Pneumocystis carinii-specific DNA probe to quantitate infection.
A repetitive genomic DNA clone (B12-2) that specifically hybridizes to Pneumocystis carinii DNA has been identified. No cross-hybridization to genomic DNA prepared from bacteria, other fungi, protozoa, or mammals was observed. Clone B12-2 is multiply represented in the P. carinii genome. By direct hybridization to DNA prepared from the lungs of immunosuppressed rats, the probe can detect the equivalent of fewer than 1,000 P. carinii organisms. A hybridization assay employing clone B12-2 has been developed to quantitate organism load in the rat model for P. carinii. Application of the assay to track the accumulation of organisms during the immunosuppression regimen as well as to monitor the efficacy of two drug therapies used clinically for the treatment of P. carinii pneumonia is described here. The clone B12-2 hybridization assay for the determination of P. carinii organism load possesses several advantageous features and thus should serve to complement conventional staining and immunohistochemical methods.
Liberator
P A
PA
Department of Genetics, Merck Research Laboratories, Rahway, New Jersey 07065.
Anderson
J W
JW
Powles
M
M
Pittarelli
L A
LA
Worley
M
M
Becker-Hapak
M
M
Graves
D C
DC
Schmatz
D M
DM
D000818:Animals; D000935:Antifungal Agents / Q000627:therapeutic use; D003001:Cloning, Molecular; D015342:DNA Probes; D004271:DNA, Fungal / Q000235:genetics; D004195:Disease Models, Animal; D008297:Male; D009693:Nucleic Acid Hybridization; D011010:Pneumocystis / Q000235:genetics* / Q000302:isolation & purification; D011020:Pneumonia, Pneumocystis / Q000188:drug therapy / Q000382:microbiology; D051381:Rats; D017207:Rats, Sprague-Dawley
D000935:Antifungal Agents; D015342:DNA Probes; D004271:DNA, Fungal