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Mark G. Rayner; Yingze Zhang, PhD; Michael C. Gorry; Yiping Chen, DDS; J. Christopher Post, MD; Garth D. Ehrlich, PhD

JAMA. 1998;279:296-299.

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.

From the Departments of Pathology (Messrs Rayner and Gorry and Drs Zhang, Chen, Post, and Ehrlich) and Otolaryngology (Drs Post and Ehrlich) and the Center for Genomic Sciences (Drs Post and Ehrlich), University of Pittsburgh School of Medicine; Department of Pediatric Otolaryngology (Drs Post and Ehrlich), Children's Hospital of Pittsburgh, Pittsburgh, Pa. Drs Zhang, Post, and Ehrlich and Mr Gorry are now with the Center for Genomic Sciences, Allegheny University of the Health Sciences, Pittsburgh, Pa. This work was performed at the Center for Genomic Sciences at the University of Pittsburgh School of Medicine.

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