Pneumolysin polymerase chain reaction for diagnosis of pneumococcal pneumonia and empyema in children

Abstract

Streptococcus pneumoniae is the most important cause of childhood pneumonia and empyema, yet the diagnosis of pneumococcal infections by conventional methods is challenging. In this study, the clinical value of the pneumolysin-targeted real-time polymerase chain reaction (PCR) method for the diagnosis of pneumococcal pneumonia and empyema was evaluated with 33 whole blood samples and 12 pleural fluid samples. The analytical sensitivity of the PCR assay was 4 fg of pneumococcal DNA, corresponding to two genome equivalents of pneumococcal DNA per reaction. The PCR assay correctly detected all clinical isolates of S. pneumoniae tested, whereas all nonpneumococcal bacterial organisms tested were negative by PCR. In a clinical trial, S. pneumoniae was detected by PCR in the pleural fluid of 75% of children with empyema, increasing the detection rate of pneumococcus almost tenfold that of pleural fluid culture. However, in whole blood samples, PCR detected S. pneumoniae in only one child with pneumonia and one child with pneumococcal empyema and failed to detect S. pneumoniae in three children with blood cultures positive for S. pneumoniae. The present data indicate that pneumolysin-targeted real-time PCR of pleural fluid is a valuable method for the etiologic diagnosis of pneumococcal empyema in children. The ease and rapidity of the LightCycler technology (Roche Diagnostics, Mannheim, Germany) make real-time PCR an applicable tool for routine diagnostics. In the evaluation of blood samples, blood culture remains the superior method for the diagnosis of bacteremic pneumococcal disease.