Polymerase chain reaction detection of Pneumocystis jiroveci: evaluation of 9 assays
0301 basic medicine
Dihydropteroate Synthase
Membrane Glycoproteins
RNA, Ribosomal, 5S
Pneumocystis carinii
DNA, Mitochondrial
DNA, Ribosomal
Polymerase Chain Reaction
Sensitivity and Specificity
3. Good health
Fungal Proteins
Pneumocystis Infections
RNA, Ribosomal, 23S
Tetrahydrofolate Dehydrogenase
03 medical and health sciences
Microscopy, Fluorescence
DNA, Ribosomal Spacer
RNA, Ribosomal, 18S
Humans
DNA, Fungal
False Negative Reactions
DOI:
10.1016/j.diagmicrobio.2007.02.014
Publication Date:
2007-08-08T11:22:54Z
AUTHORS (3)
ABSTRACT
Various polymerase chain reaction (PCR) amplification strategies have been described for detecting Pneumocystis jiroveci in clinical specimens. Different combinations of primer/target and platforms have been reported to yield varying PCR detection rates. PCR was evaluated on clinical specimens using internal transcribed spacer regions of the rRNA nested, dihydropteroate synthase single and nested, dihydrofolate reductase nested, major surface glycoprotein heminested, mitochondrial large subunit rRNA (mtLSUrRNA) single and nested, 18S rRNA 1-tube nested, and real-time 5S rRNA PCR. The most sensitive PCR was subsequently compared with routine diagnostic immunofluorescence (IF) microscopy. Discrepant PCR and IF results were resolved after review of clinical and histology/cytology records. Major discrepancies were observed among the methods investigated. mtLSUrRNA nested PCR was the most sensitive, produced less false-negative results, and displayed the highest degree of concordance with histology. Direct comparison of mtLSUrRNA nested PCR versus IF yielded low sensitivity and specificity, which were improved for PCR and lowered for IF on review of clinical and laboratory records.
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