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J Med Microbiol 57 (2008), 1205-1212; DOI: 10.1099/jmm.0.2008/000935-0
© 2008 Society for General Microbiology
ISSN 1473-5644

Molecular detection methods and serotyping performed directly on clinical samples improve diagnostic sensitivity and reveal increased incidence of invasive disease by Streptococcus pneumoniae in Italian children

Chiara Azzari, Maria Moriondo, Giuseppe Indolfi, Cristina Massai, Laura Becciolini, Maurizio de Martino and Massimo Resti

Department of Paediatrics, University of Florence and Anna Meyer Children’s Hospital, Viale Pieraccini 24, Florence I-50139, Italy

Correspondence
Chiara Azzari
chiara.azzari{at}unifi.it

Received 2 February 2008
Accepted 22 April 2008

The aims of this study were to evaluate the incidence of invasive pneumococcal disease (IPD) in Italian children and perform serotyping by PCR-based assays directly on clinical samples. A 1-year paediatric (0–14 years) population-based surveillance study was designed to evaluate the incidence of IPD in the province of Florence, Italy, by cultural and molecular methods. Among 92 children (80 with pneumonia, 8 with meningitis/sepsis, 4 with arthritis), 4 cases of IPD were diagnosed both by culture and real-time PCR and 18 cases exclusively by molecular methods. The sensitivity of molecular methods was significantly higher than that of cultural methods (Cohen’s {kappa} 0.41; McNemar P=0.000008). The incidence of IPD in children below 2 years of age was 11.5/100 000 and 51.8/100 000 by cultural and molecular methods, respectively. Pneumococcal serotyping by multiplex sequential PCR was obtained in 19/22 samples. Real-time PCR and multiplex sequential PCR can be used directly on biological samples, improving the ability to diagnose IPD. The incidence of IPD appears 5–10 times higher by PCR than by cultural methods.

Abbreviations: CSF, cerebrospinal fluid; IPD, invasive pneumococcal disease; RT, real-time.




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