This program was
repeated for 35 cycles. In every run, both positive and negative controls were considered. PCR Product Detection Two percent agarose gel electrophoresis was used. Twenty five µl of the PCR products were mixed with two µl of 6x loading buffer dye and loaded into the individual wells. The electrophoresis was performed in Tris Acetate EDTA (TAE) buffer for one hour. At the end, the gel was stained in ethidium bromide solution(1 µg/ml) for 15 minutes. The results were analyzed according to the product length which Inhibitors,research,lifescience,medical were visualized on gel documentation system and photographed. Data were analyzed using Statistical Package for Social Sciences (SPSS, version 14). The results of the bacterial cultures and PCR assays were analyzed using Chi-Square test. Differences between the groups were considered statistically significant if P values were <0.05. Results A total of 36 patients including 23 boys and 13 girls with a mean±SD age of 6.72±2.95 Inhibitors,research,lifescience,medical years (range; 2-13 years) participated in the study. Twenty
seven (75%) had bilateral and nine (25%) had unilateral OME, therefore, a total of 63 samples were obtained. Two patients were identified incidentally during routine examination of ear, nose and throat, and remaining 34 patients presented with chief complaints of hearing impairment (70%), otalgia (24%), or both (6%). The mean duration of symptoms Inhibitors,research,lifescience,medical (hearing loss and/or otalgia) was 6 months (Range; 2-14). One patient had a history of previous tympanostomy tube insertion. Glue was the most common (n=50, 79.4%) type of aspirated fluid. Ten (15.9%) of ears had serous fluid. Purulent material was seen only in three (4.8%) of the ears. The results of PCR and Inhibitors,research,lifescience,medical bacterial culture are presented in table 1 and and2.2. In the standard Olaparib bacteriologic culture, bacterial growth was detected in 38 (60.3%) samples. The most frequent pathogens Inhibitors,research,lifescience,medical were S. pneumoniae, H. influenzae, M .catarrhalis and coagulase negative Staphylococci (49.2%). The percentages of culture positive effusions for
S. pneumoniae, H. influenzae and M. catarrhalis were 15.9%, 9.5%, and 9.5%, respectively. PCR assay was done for because three of frequently-occuring bacterial pathogen(s) in OME including S. pneumoniae, H. influenzae and M. catarrhalis. The DNAs of one or more of these bacteria were present in 60 (95.2%) samples. The DNAs of S. pneumoniae, M. catarrhalis and H. influenzaee were detected in 19%, 36.5% and 95.2% of the samples, respectively. In 32 (50.7%) samples, the DNA of only one bacterial species, and in 28 (44.5%) samples the DNAs of more than one bacterial species were detected. Three (4.8%) samples had no DNA content. Also, the number of H. influenzae isolate was significantly higher than those for other bacteria (P<0.05). The representative results of multiplex PCR are displayed in figure 1.