Real time quantitative PCR and RT--PCR for analysis of Pneumocystis carinii hominis.
Pneumocystis carinii hominis is a common cause of pneumonia in immunocompromised patients and particularly in those infected by HIV. Giemsa- and Gomori--Grocott-stained smears are widely used for detection and quantification of this opportunistic fungus obtained from biological samples or from in vitro culture. But these methods are fastidious and time-consuming. Thus, instead of performing a count of organisms, we focused our attention on the level of specific DNA by a quantitative PCR technique. This procedure has the advantage of greater precision and more objectivity. To verify the presence of organisms, quantitative RT--PCR based on DHFR and a cell cycle mRNA have been developed. In this current study, we present a detailed description of these methods and their applications for analysis of P. carinii hominis.
Kaiser
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Laboratoire de Parasitologie, Mycologie Médicale et Maladies Tropicales--Hospices Civils de Lyon-EA 3087-Université C. Bernard Lyon I-8, avenue Rockefeller 69373, Lyon Cedex 08, France. kaiser@rockefeller.univ-lyon1.fr
Rabodonirina
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Picot
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D000818:Animals; D052160:Benzothiazoles; D016203:CDC2 Protein Kinase / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003959:Diamines; D005456:Fluorescent Dyes / Q000737:chemistry; D018816:Genes, cdc; D006801:Humans; D008856:Microscopy, Fluorescence; D009930:Organic Chemicals; D011010:Pneumocystis / Q000235:genetics* / Q000302:isolation & purification; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis / Q000382:microbiology*; D011804:Quinolines; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry / Q000235:genetics
D052160:Benzothiazoles; D004271:DNA, Fungal; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D013762:Tetrahydrofolate Dehydrogenase; D016203:CDC2 Protein Kinase