COMPARISON OF PCR ASSAY WITH BACTERIAL CULTURE FOR DETECTING STREPTOCOCCUS-PNEUMONIAE IN MIDDLE-EAR FLUID OF CHILDREN WITH ACUTE OTITIS-MEDIA

We have studied etiological diagnosis of acute otitis media (AOM) by c omparing a newly developed pneumococcal PCR for Streptococcus pneumoni ae to bacterial culture with 180 middle ear fluid (MEF) samples of 125 children with 125 episodes of AOM. For pneumococcal PCR assay, DNA fr om MEF samples was extracted by phenol-chloroform. The outer primers u sed amplified a 348 bp region of the pneumolysin gene, and the inner p rimers amplified a 208-bp region. S. pneumoniae was cultured in 33 (18 %) samples, and pneumolysin PCR was positive for 51 (28%) of 180 MEF s amples. Only 2 of 21 PCR-positive, S. pneumoniae culture-negative samp les were positive for other otitis pathogens. By combining MEF culture and PCR results, 54 (30%) of 180 MEF samples had evidence of pneumoco ccal etiology. In conclusion, pneumolysin PCR is a sensitive and speci fic new method to study pneumococcal involvement in MEF samples of chi ldren with AOM.