Quantification of the detection of Pneumocystis carinii by DNA amplification.
We have developed a highly specific and sensitive technique for the detection of Pneumocystis carinii DNA using DNA amplification by the polymerase chain reaction (PCR). PCR products are detected by agarose gel electrophoresis and Southern hybridization to an oligonucleotide probe. Here we report the calibration of parasite numbers with amplification and hybridization signals and show that we can detect P. carinii to a lower limit of one to two organisms. The quantification of this diagnostic technique allows us to establish the number of organisms in a clinical sample which correspond to pneumocystis pneumonia or to sub-clinical pulmonary colonization.
Peters
S E
SE
Department of Paediatrics, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, UK.
Wakefield
A E
AE
Banerji
S
S
Hopkin
J M
JM
D000818:Animals; D004271:DNA, Fungal / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008168:Lung / Q000382:microbiology; D011010:Pneumocystis / Q000302:isolation & purification*; D016720:Pneumocystis Infections / Q000382:microbiology; D016133:Polymerase Chain Reaction; D051381:Rats; D011919:Rats, Inbred Strains
D004271:DNA, Fungal