Evidence of bacterial metabolic activity in culture-negative otitis media with effusion

Context.-Otitis media with effusion (OME) can lead to significant hearing loss in children. Although previous studies have shown that bacterial DNA is present in a significant percentage of effusions sterile by culture, whether the DNA represents viable organisms or "fossilized remains" is unknown. Objective.-To determine if bacterial messenger RNA (mRNA), as detected by a reverse transcriptase-polymerase chain reaction (RT-PCR)-based assay, is present in chronic pediatric middle ear effusions that contain bacterial DNA but are sterile by standard cultural methods. Bacterial mRNAs have a half-life measured in seconds to minutes; therefore, detection of bacteria-specific mRNAs would be evidence that metabolically active organisms are present. Design.-Blinded comparative study. Patients.-A total of 93 effusions from pediatric outpatients seen for myringotomy and tube placement for chronic (>3 months) OME (median age of children, 17 months). Setting.-Tertiary care pediatric hospital. Main Outcome Measures.-Percentage of positive test results for RT-PCR-based assays compared with culture for Haemophilus influenzae and concordance between RT-PCR and PCR-based findings for bacterial nucleic acids. Results.-Eleven (11.8%) of the 93 specimens tested positive by culture, PCR, and RT-PCR for H influenzae, A total of 29 specimens (31.2%) were positive by PCR but negative by culture for H influenzae. All 29 specimens were positive by RT-PCR for H influenzae-specific mRNA. Conclusions.-The RT-PCR-based assay system can detect the presence of bacterial mRNA in a significant percentage of culturally sterile middle ear effusions, establishing the presence of viable, metabolically active, intact organisms in some culture-negative OME.