DEVELOPMENT AND EVALUATION OF A RAPID AND SIMPLE PROCEDURE FOR DETECTION OF PNEUMOCYSTIS-CARINII BP PCR

We report the development of a simplified PCR-based assay for the dete ction of Pneumocystis carinii DNA in clinical specimens. The adoption of a rapid DNA extraction procedure and the introduction of a type of enzyme-linked immunosorbent assay for PCR product detection enabled th is procedure to be carried out in a single working day in a clinical m icrobiology laboratory. The PCR assay was prospectively compared with an immunofluorescent antibody (FA) staining method for the detection o f P. carinii in induced sputum and bronchoalveolar lavage (BAL) specim ens. The results of the study showed that, for induced sputum specimen s, FA staining had a sensitivity of 78% (32 of 41 specimens) and a spe cificity of 100% (166 of 166 specimens); PCR was 100% (41 of 41 specim ens) sensitive and 98% (162 of 166 specimens) specific. For BAL specim ens, FA staining was 100% sensitive (21 of 21 specimens) and 100% spec ific (133 of 133 specimens), and PCR had a sensitivity of 100% (21 of 21 specimens) and a specificity of 99% (132 of 133 specimens). These r esults strongly suggest that use of our PCR-based assay could effect c linically useful improvements in the sensitivity of induced sputum spe cimens for the detection of P. carinii.