Total DNA from the adenoid specimens (7 RAOM and 13 OSA) were purified using DNEasy Blood and Tissue Kit (Qiagen, Valencia, CA), following the manufacturerfs protocol for DNA isolation in Gram-positive bacteria, with modifications.
Power analysis was conducted using G*Power statistical analysis, and using the criteria that at least 25% difference in a species abundance between RAOM and OSA will be observed (based on a previous similar study), a 20% standard deviation in each group, an [alpha] of 0.05 and power of 0.80, we determined that at least 7 samples is needed for each group for the pyrosequencing analysis.
salivarius qPCR were normalized with the 16S threshold cycle ([C.sub.t]) and were compared between the OSA and RAOM groups using t-test (JMP.
Adenoid specimens were obtained from 13 patients with OSA and 7 patients with RAOM. The ages of the subjects ranged from two to 11 years, with a mean age of 5.6 years (Table 1).
All adenoid specimens analyzed with 454 pyrosequencing (n=5 each for OSA and RAOM) had evidence of microbes (Figure 1).
salivarius, we found that the threshold cycle (Ct; the number of cycles required for the fluorescent signal to cross a threshold; i.e., exceeds background level, and is inversely proportional to the amount of target DNA in the sample) of 16S was not different between the OSA and RAOM group (Figure 2a).
As differences at the species level may be clinically significant, the goal of the present study was to compare the microbiome of adenoids, at the species level, from patients with RAOM or OSA .
catarrhalis tend to be higher in the RAOM group, though the differences were not statistically significant.
In the present study, the relative abundance of Streptococcus salivarius was significantly higher in the OSA compared to the RAOM group.
The use of TT for rAOM in children who do not have associated OME does not seem to be supported by the current available evidence, but children with rAOM and associated OME have been shown to derive short-term (<6 months) benefit from TT placement.
Caution must be exercised when interpreting the available evidence and when a clinician considers withholding TT placement for rAOM, because high-quality studies that can be generalized to current otolaryngology practice do not exist, and QOL studies examining the effects of TT placement in rAOM are also lacking.