Abstract
The diagnosis of Pneumocystis jirovecii pneumonia is based on observation of the microorganism using several staining techniques in respiratory samples, especially bronchoalveolar lavage and induced sputum. Recently, the fungus also has been detected in oropharyngeal wash samples, but only using molecular tests. The diagnostic yield of two microscopic stains, toluidine blue O and direct fluorescent antibody, was compared in bronchoalveolar lavage and oropharyngeal wash samples for the detection of P. jirovecii in immunocompromised patients with pneumonia. Cross-sectional evaluation diagnostic tests were used in 166 immunosuppressed patients with suspected P. jirovecii. By protocol, bronchoscopic bronchoalveolar lavage and oropharyngeal wash samples were prepared by cytocentrifugation, and slides were stained with toluidine blue and fluorescent antibody. The proportion of positive results from each stain and concordance between them were determined. Twenty-four cases (14.5%) of P. jirovecii were detected in bronchoalveolar lavage samples. Of them, 21 were positive by both toluidine blue and fluorescent antibody stains, whereas 3 cases were detected by fluorescent antibody alone. None of the 166 oropharyngeal wash samples were positive by either of these techniques. No significant differences were found between proportions from positive results (p=0.63). Concordance (kappa coefficient) between both stains was 0.92 (95% CI: 0.84-1.00). Both techniques were useful to diagnose P. jirovecii in bronchoalveolar lavage samples. However, toluidine blue stain did not detect 12% of fluorescent antibody positive cases. Oropharyngeal wash samples do not provide sufficient material for the microscopic identification of this fungus.
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