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

Holder RC, Kirse DJ, Evans AK, Peters TR, Poehling KA, Swords WE, Reid SD - BMC Pediatr (2012)

Bottom Line: Haemophilus influenzae was the predominant single organism and caused 58% of all AOM in this study.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology and Immunology, Wake Forest School of Medicine, Winston Salem, NC 27101, USA.

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.

Show MeSH

Related in: MedlinePlus

Multiplex PCR Results. Multiplex PCR products were electrophoresed on 2.5% agarose. Hi, Haemophilus influenzae control band, expected size 525 bp; Sp, Streptococcus pneumoniae control band, expected size 484 bp; Ao, Alloiococcus otitidis control band, expected size 264 bp; Mc, Moraxella catarrhalis control band, expected size 237 bp; 265L, 269R, 246R, and 248L, experimental samples; bp, base pair.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3475091&req=5

Figure 1: Multiplex PCR Results. Multiplex PCR products were electrophoresed on 2.5% agarose. Hi, Haemophilus influenzae control band, expected size 525 bp; Sp, Streptococcus pneumoniae control band, expected size 484 bp; Ao, Alloiococcus otitidis control band, expected size 264 bp; Mc, Moraxella catarrhalis control band, expected size 237 bp; 265L, 269R, 246R, and 248L, experimental samples; bp, base pair.

Mentions: A multiplex PCR procedure was performed to simultaneously detect H. influenzae, S. pneumoniae, A. otitidis, and M. catarrhalis with minor modifications of the methods described by Hendolin et al. [20]. Briefly, PCR extension steps were performed at 65°C with HotMasterMix (5 Prime, Gaithersburg, MD) and using the primers listed in Table 1. H. influenzae 86–028, S. pneumoniae TIGR4, A. otitidis SS1337, and M. catarrhalis 7169 were used as positive controls for PCR (Figure 1). Results were visualized using agarose gel electrophoresis (2.5% agarose, 100 V for 3 h) and ethidium bromide staining (500 ng/mL final ethidium bromide concentration).


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

Holder RC, Kirse DJ, Evans AK, Peters TR, Poehling KA, Swords WE, Reid SD - BMC Pediatr (2012)

Multiplex PCR Results. Multiplex PCR products were electrophoresed on 2.5% agarose. Hi, Haemophilus influenzae control band, expected size 525 bp; Sp, Streptococcus pneumoniae control band, expected size 484 bp; Ao, Alloiococcus otitidis control band, expected size 264 bp; Mc, Moraxella catarrhalis control band, expected size 237 bp; 265L, 269R, 246R, and 248L, experimental samples; bp, base pair.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3475091&req=5

Figure 1: Multiplex PCR Results. Multiplex PCR products were electrophoresed on 2.5% agarose. Hi, Haemophilus influenzae control band, expected size 525 bp; Sp, Streptococcus pneumoniae control band, expected size 484 bp; Ao, Alloiococcus otitidis control band, expected size 264 bp; Mc, Moraxella catarrhalis control band, expected size 237 bp; 265L, 269R, 246R, and 248L, experimental samples; bp, base pair.
Mentions: A multiplex PCR procedure was performed to simultaneously detect H. influenzae, S. pneumoniae, A. otitidis, and M. catarrhalis with minor modifications of the methods described by Hendolin et al. [20]. Briefly, PCR extension steps were performed at 65°C with HotMasterMix (5 Prime, Gaithersburg, MD) and using the primers listed in Table 1. H. influenzae 86–028, S. pneumoniae TIGR4, A. otitidis SS1337, and M. catarrhalis 7169 were used as positive controls for PCR (Figure 1). Results were visualized using agarose gel electrophoresis (2.5% agarose, 100 V for 3 h) and ethidium bromide staining (500 ng/mL final ethidium bromide concentration).

Bottom Line: Haemophilus influenzae was the predominant single organism and caused 58% of all AOM in this study.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology and Immunology, Wake Forest School of Medicine, Winston Salem, NC 27101, USA.

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.

Show MeSH
Related in: MedlinePlus