Molecular Diagnosis of Streptococcus pneumoniae in Middle Ear Fluids from Children with Otitis Media with Effusion

PURPOSE: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. METHODS: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. RESULTS: The detection limit of the PCR assay was approximately 10(4) colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over 10(6) CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). CONCLUSIONS: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA-specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.

Detection of Fungus and Bacteria in Otitis Media with Effusion of Children Using Polymerase Chain Reaction and Its Correlation of Clinical Factors / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Several hypotheses have been proposed in the etiopathogenesis of otitis media with effusion (OME). The bacterial or fungal infection may also play a major role in the pathogenesis of OME. To investigate the relationship between bacteria and fungi as pathogens in OME and to analyze the incidence, the authors evaluated the pathogens of OME using polymerase chain reaction (PCR) technique, which is known to be more sensitive and specific than conventional bacterial and fungal culture. SUBJECTS AND METHOD: Twenty-three children (thirty ears), who were diagnosed with OME and had underwent ventilation tube insertion, were evaluated in the department of ORL-HNS, University Hospital, from May 2006 to March 2007. The middle ear effusion, obtained during the procedure, was evaluated for the identification of bacteria and fungi by PCR. RESULTS: Among 30 ears, viable pathogenic bacteria were detected in 20 ears (66.6%) and fungi in 13 ears (43.3%). The bacterial pathogens included Haemophilus influenzae (13 ears), followed by Streptococcus pneumonize (6 ears). The fungi detected from PCR were Candida albicans (4 ears-30.8%), Aspergillus niger (2 ears-15.4%) and Paecilomyces lilacinus (2 ears-15.4%). Additional pathogens include Basidiomycete yeast, Saccharamycete sp., Eurotium rubrum, Dothioraceae sp. and Stemphylium solani. Detection of fungal DNA was more common in patients with cleft palate and in recurrent cases with statistical significance. CONCLUSION: The use of PCR of middle ear effusion is effective for the detection of pathogens in patients with OME. While bacteria were thought to be the causative pathogen, this study suggests the etiological role of fungi in the pathogenesis of OME. However, the relationship between fungi and OME requires further studies.

The Expression and Significance of Transforming Growth Factor ?1 in the Middle Ear Mucosa of Guinea Pigs with Otitis Media With Efussion / 听力学及言语疾病杂志

Objective To explore the expression of transforming growth factor ?1(TGF-?1)in the mucosa of guinea pigs with otitis media with effusion(OME)and the role of TGF-?1 in the development of OME.Methods 70 guinea pigs were randomly devided into 7 groups,each with 10 animals.OME was produced by injecting deactivated streptococcus pneumoniae into the typmpanic cavity of guinea pigs.The expression of TGF-?1 was examined by means of immunohistochemistry 6 h,1 d,3 d,7 d,14 d and 30 d after injection.Results TGF-?1 expression could be detected in the middle ear mucosa at 7 d after injection and reached maximum at 30 d.Conclusion TGF-?1 is expressed in the middle ear mucosa of the guinea pigs with OME induced by deactivated streptococcus pneumoniae,and this indicates that TGF-?1 may play a role in the chronic protraction of OME.

Compositional difference of middle ear effusion according to viscosity of the effusion / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Serous otitis media (SOM) is usually responsive to medical treatment, whereas mucoid otitis media (MOM) is not. This study was undertaken to elucidate the compositional difference between serous and mucoid effusion, and to investigate whether MUC5AC acts as a major mucin in the middle ear mucosa with MOM. MATERIALS AND METHODS: Middle ear effusion (MEE) samples were collected from 29 patients with MOM and 20 patients with SOM. The level of mucin, lysozyme, secretory IgA and interleukin-8 were measured by dot blotting or enzyme-linked immunosorbent assay. Periodic acid-Schiff and immunohistochemical staining with monoclonal anti-MUC5AC antibody were performed on the serial sections of middle ear mucosa with MOM. RESULTS: Mucoid effusions contained higher levels of mucin, lysozyme, secretory IgA and interleukin-8 than serous effusion. Immunohistological study revealed that MUC5AC mucin was expressed in only a small portion of the goblet cells of middle ear mucosa with MOM. CONCLUSION: Our study suggests that both serous secretions and mucin might make the middle ear effusion more viscous, and that mucins other than MUC5AC might have a major role in the viscosity of MEE. Further study is necessary to identify the major mucins in the MEE of otitis media with effusion.

The Prevalence of Otitis Media with Effusion among Kindergarten and Elementary School Children in Song Buk, Seoul, Korea and Risk Factors / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Otitis media with effusion (OME) is the most common bacterial infection and the most frequent otologic indication for antimicrobial or surgical therapy in young children. It is also the leading cause of hearing disturbance in children. There have been few reports about the prevalence rate of OME in preschool children in Seoul. So, we investigated the prevalence of OME among kindergarten and elementary children in Song Buk Gu, Seoul, Korea. MATERIAL AND METHODS: Tympanometry and pneumatic otoscopic examination were performed on 3,364 children aged 3-7 years, from March to June of 1999. RESULTS: A total of 11.74% of the subjects had unilateral or bilateral OME. The prevalence rate of OME in 3 year-old children was 30.0% and that of 7 year-old children was decreased to 8.9%. The prevalence rate in March was 18.5% and that in June decreased to 6.5%. The majority of the cases were free of subjective symptoms, but some of significant symptoms were ear fullness, otalgia, tendency to increase the TV volume and autophonia. CONCLUSION: Significant risk factors are seasonal variation, age and the history of OME. There should be more attention paid to OME in children aged 7 years and younger, because its high prevalence and less symptoms.

The Application and Characteristic of Auditory Brainstem Response in Children with Otitis Media with Effusion / 听力学及言语疾病杂志

Objective To explore the characteristics of auditory brainstem response(ABR) in children with otitis media with effusion(OME) before and after myringotomy with grommet insertion and the significance of ABR in diagnosing OME.Methods ABR were recorded in 50 cases(100 ears) before receiving myringotomy with grommet insertion for OME,30 cases(60 ears) underwent ABR monitoring after surgery again.For comparison,50 age-matched healthy children(100 ears) were also tested with ABR.Furthermore,30 cases were divided into two groups according to the viscidity of tympanic cavity secretion and the response threshold of wave Ⅴ were compared between the two groups.Results The response threshold of wave Ⅴand the peak latency of wave Ⅰwere normal in 13 percent of 50 cases(100 ears) before surgery.The rate of missed diagnosis was 13%.Among 50 cases,41% had normal response threshold for wave Ⅴ,52% had slight abnormal responses of thresholds of wave Ⅴ and 7% had medium response threshold of wave Ⅴ.The peak latency of wavs Ⅰwas normal in 19% OME children,but it prolonged in 72% OME children and absent in 9% OME children.The prolonged peak latencies(waves Ⅰ,Ⅲ andⅤ) and eleveted response thresholds of wave Ⅴ and shorted transmission time between waves(Ⅰ-Ⅲ andⅠ-Ⅴ) significant when compared to the control group.After surgery,the response thresholds of wave Ⅴand the peak latency of wave Ⅰwere normal in 46.7% children,the response threshold of wave Ⅴ were normal in 70.5% and slight abnormal in 29.5%.Occuring(elicit) rate of wave Ⅰwas 100% and peak latency of wave Ⅰwas normal in 50.2% chilren.The differrence of peak latencies(waves Ⅰ,Ⅲ andⅤ) and the response threshold of wave Ⅴ before surgery

Rat Model of Otitis Media with Effusion / 听力学及言语疾病杂志

Objective To establish an otitis media with effusion model in rat,and provide a pure physical approach of animal model for researching the otitis media with effusion.Methods Twenty-four clean grade male SD rats were selected.Only the right ears were used for the model group,the contralateral ears were used as the control group.The animal model of otitis media with effusion was established by blocking eustachian tube with cork stopper through the right side mandible,the hearing was evaluated by ABR and tympanogram configurations,the pathologic changes of middle ear and eustachian tube mucous membrane were examined by light microscopy.Results Twenty out of 24 ears of rats' of hearing threshold increased from 34.25?5.45 dB to 57.63?5.46 dB (P