One third of middle ear effusions from children undergoing tympanostomy tube placement had multiple bacterial pathogens

Abstract

Background Because previous studies have indicated that otitis media may be a polymicrobial disease, we prospectively analyzed middle ear effusions of children undergoing tympanostomy tube placement with multiplex polymerase chain reaction for four otopathogens. Methods Middle ear effusions from 207 children undergoing routine tympanostomy tube placement were collected and were classified by the surgeon as acute otitis media (AOM) for purulent effusions and as otitis media with effusion (OME) for non-purulent effusions. DNA was isolated from these samples and analyzed with multiplex polymerase chain reaction for Haemophilus influenzae , Streptococcus pneumoniae , Alloiococcus otitidis , and Moraxella catarrhalis . Results 119 (57%) of 207 patients were PCR positive for at least one of these four organisms. 36 (30%) of the positive samples indicated the presence of more than one bacterial species. Patient samples were further separated into 2 groups based on clinical presentation at the time of surgery. Samples were categorized as acute otitis media (AOM) if pus was observed behind the tympanic membrane. If no pus was present, samples were categorized as otitis media with effusion (OME). Bacteria were identified in most of the children with AOM (87%) and half the children with OME (51%, p < 0.001). A single bacterial organism was detected in middle ear effusions from children with AOM more often than those with OME (74% versus 33%, p < 0.001). Haemophilus influenzae was the predominant single organism and caused 58% of all AOM in this study. Alloiococcus otitidis and Moraxella catarrhalis were more frequently identified in middle ear effusions than Streptococcus pneumoniae . Conclusions Haemophilus influenzae , Streptococcus pneumoniae , Alloiococcus otitidis , and Moraxella catarrhalis were identified in the middle ear effusions of some patients with otitis media. Overall, we found AOM is predominantly a single organism infection and most commonly from Haemophilus influenzae . In contrast, OME infections had a more equal distribution of single organisms, polymicrobial entities, and non-bacterial agents.

Moraxella catarrhalis Might Be More Common than Expected in Acute Otitis Media in Young Finnish Children

“…In these treatment groups, M. catarrhalis was detected in 14, 11, 15, and 2 episodes, a This instance of PCR negativity of a sample containing H. influenzae was a consequence of the sequence of the forward primer. Some strains of H. influenzae are not detectable by the primer used in this study (16,24,32). The presence of H. influenzae in a high quantity was verified by sequencing the V3-to-V5 region of the bacterial 16S rRNA gene using generic primers.…”

“…To reinforce the finding, and to amend the design of the detection primer, we then retrieved available sequences of the 16S region of H. influenzae from GenBank, aligned them, removed the ambiguous sequences, and assessed the primer annealing site in the 557 remaining sequences. The H. influenzae-specific forward primer designed by Holder et al (16) and used here annealed with no mismatches to 401 of 557 retrieved sequences. Two other prevalent motifs were related to the forward primer annealing site: the first one differed by only two nucleotides in the 5=-proximal portion of the primer, so safe detection by the original primer may be reasonably expected; it was present in 53 of 557 retrieved sequences.…”

“…The primers and probes used are summarized in Table 1. Four assays were targeted to the 16S rRNA genes (an H. influenzae assay, an assay for S. pneumoniae, S. oralis, S. mitis, and S. infantis, an A. otitidis assay, and an M. catarrhalis assay) with primers described by Holder et al (16) complemented with a hydrolysis probe described by Nadkarni et al (17) which allowed adaptation of the assay to the real-time PCR format. Every PCR product of the 16S-based assays was verified by bidirectional sequencing using BigDye Terminator 3.1 chemistry on an ABI 3130xl capillary sequencer (Life Technologies, Foster City, CA).…”

Source of bacterial RNA in chronic otitis media with effusion

“…PCR is a sensitive method for detecting bacterial DNA, requiring only small DNA fragments for amplification. It is thought that detection of bacterial DNA by PCR only represents the presence of live bacteria (Post et al, 1996;Hendolin et al, 2000;Holder et al, 2012). Studies involving PCR have shown that bacterial DNA exists in some MEE samples.…”

The Vaccine Candidate Substrate Binding Protein SBP2 Plays a Key Role in Arginine Uptake, Which Is Required for Growth of Moraxella catarrhalis

“…M. catarrhalis is a common cause of acute otitis media in children, along with nontypeable Haemophilus influenzae and Streptococcus pneumoniae (6). Studies using PCR to detect bacterial DNA in middle ear fluid reveal that M. catarrhalis plays an even larger role in otitis media than revealed by culture alone (15)(16)(17)(18). With the increasing widespread administration of pneumococcal conjugate vaccines globally, patterns of nasopharyngeal colonization in children and the distribution of pathogens causing otitis media are undergoing changes, with M. catarrhalis taking on a more prominent role (19).…”