Biomolecular Techniques to Detect Pneumocystis carinii f. sp. hominis Pneumonia in Patients with
Acquired Immunodeficiency Syndrome
Chiara Atzori, MD; Elena Angeli, MD; Fiorenza Agostoni, MD; Annalisa Mainini, MD; Valeria Micheli,
MD; and Antonietta Cargnel, MD
Int J Infect Dis 1999; 3:76-81.
Objectives: To verify the clinical value of two different polymerase chain reactions (PCRs) for
noninvasive diagnosis and follow-up during Pneumocystis carinii f. sp. hominis pneumonia (PCP) and to
analyze the P. carinii f. sp. hominis genotypes involved. Methods: Internal transcribed spacers (ITSs)
nested PCR was applied to 630 samples (bronchoalveolar lavage, sera, peripheral blood mononuclear
cells, and oropharyngeal samples) from 122 patients with acquired immunodeficiency syndrome and
pneumonia and 40 control samples from 20 subjects seronegative for human immunodeficiency virus.
One hundred and eighty samples also were examined by mt-rRNA PCR. Bronchoalveolar lavage
samples and 33 sera were analyzed by type-specific oligonucleotide hybridization. Results: On
bronchoalveolar lavage samples, the two PCRs consistently confirmed the morphologic diagnosis of
PCP. The sensitivity of ITSs nested PCR versus mt-rRNA PCR was 57.3% versus 14.3% on sera,
32.3% versus 22.8% on peripheral blood mononuclear cells, and 69.1% versus 48.6% on
oropharyngeal samples (garglings). Both PCRs had 100% specificity. Type-specific oligonucleotide
hybridization revealed in 72.2% of bronchoalveolar lavage samples a single P. carinii f. sp. hominis
genotype, whereas in 27.8% co-infection with more than one strain was detected. Conclusion: On
noninvasive samples, ITSs nested PCR was more sensitive than mt-rRNA PCR, and it confirmed the
diagnosis in all patients with PCP. For each patient with PCP at least one noninvasive sample was
positive for P. carinii f. sp. hominis DNA.
KEYWORDS:P. carinii. f sp. hominis, PCR, typing
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