PCR ASSAY FOR DETECTING STREPTOCOCCUS-PNEUMONIAE IN THE MIDDLE-EAR OFCHILDREN WITH OTITIS-MEDIA WITH EFFUSION

Detecting We compared a newly developed pneumococcal polymerase chain reaction (PCR) for Streptococcus pneumoniae (Pnc) to bacterial culture in 123 middle ear effusion (MEE) samples of 123 children with otitis media with effusion (OME). For the pneumococcal PCR assay, DNA of MEE samples was purified by a QIAamp blood kit. The outer primers used amp lified a 348 basepair region of the pneumolysin gene, and the inner a 208. Pnc was cultured in 14 (11%) and pneumolysin PCR was positive in 57 (46%) of the 123 MEE samples. All the culture positive samples were also PCR-positive. Both the samples with culturable Pnc and with posi tive pneumolysin PCR increased with shorter duration of OME and a grea ter number of acute otitis media during the preceding 6 months. In con clusion, pneumolysin PCR suggests pneumococcal involvement in MEE even in OMEs with no evidence of Pnc in culture, and thus offers a good di agnostic tool when a more accurate and sensitive pneumococcal diagnosi s is needed.