Development of a rapid real-time PCR assay for quantitation of Pneumocystis carinii f. sp. carinii.

ABSTRACT: A method for reliable quantification of Pneumocystis carinii in research models of P. carinii pneumonia (PCP) that is more convenient and reproducible than microscopic enumeration of organisms would greatly facilitate investigations of this organism. We developed a rapid quantitative touchdown (QTD) PCR assay for detecting P. carinii f. sp. carinii, the subspecies of P. carinii commonly used in research models of PCP. The assay was based on the single-copy dihydrofolate reductase gene and was able to detect <5 copies of a plasmid standard per tube. It was reproducibly quantitative (r = 0.99) over 6 log values for standards containing > or =5 copies/tube. Application of the assay to a series of 10-fold dilutions of P. carinii organisms isolated from rat lung demonstrated that it was reprodu

SUBMITTER: Larsen HH 

PROVIDER: S-EPMC120631 | biostudies-literature | 2002 Aug

REPOSITORIES: biostudies-literature