Direct detection of bacterial Biofilms on the middle-ear mucosa of children with chronic otitis media
Direct detection of bacterial Biofilms on the middle-ear mucosa of children with chronic otitis media
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DOI：
10.1001/jama.296.2.202
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发表时间：
2006-07-12
影响因子：
120.7
通讯作者：
Kerschner, Joseph E.
中科院分区：
文献类型：
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作者：
Hall-Stoodley, Luanne;Hu, Fen Ze;Kerschner, Joseph E.
背景慢性中耳炎（OM）是一种常见的儿科感染性疾病。先前的研究表明，代谢活性细菌存在于培养阴性的小儿中耳积液中，并且在灰鼠中耳炎模型中流感嗜血杆菌的实验性感染导致粘附的粘膜生物膜的形成，这表明慢性OM可能是由粘膜生物膜感染引起的。从26名儿童中获得中耳粘膜（MEM）活检标本（平均年龄，2.5 [范围，0.5-14]岁），接受鼓膜造口置管治疗渗出性中耳炎（OME）和复发性OM，并使用微生物培养、基于聚合酶链反应（PCR）的诊断进行分析，直接显微镜检查、荧光原位杂交和免疫染色。未感染（对照）MEM标本来自接受人工耳蜗植入的3名儿童和5名成人。患者在2004年2月和2005年4月之间从一个单一的美国三级转诊耳鼻喉科practice.Main结果措施共聚焦激光扫描显微镜（CLSM）图像获得MEM活检标本，并进行了生物膜形态使用通用染料和物种特异性探针流感嗜血杆菌，肺炎链球菌，卡他莫拉菌进行评价。结果在26例接受鼓膜置管的患儿中，13例（50%）有中耳炎，20例（77%）有复发性中耳炎，7例（27%）有两种诊断; 27/52（52%）耳有积液，24/24例积液至少1种OM病原体PCR阳性，6/27例积液任何病原体培养阳性。使用通用和病原体特异性探针，CLSM在来自OME和复发性OM儿童的50份MEM标本中的46份（92%）上观察到粘液生物膜。生物膜没有观察到8个控制MEM标本从接受人工耳蜗implantation.Conclusion MEM活检标本从OME和复发OM的儿童生物膜的直接检测支持的假设，这些慢性中耳疾病是生物膜相关的。
Context Chronic otitis media ( OM) is a common pediatric infectious disease. Previous studies demonstrating that metabolically active bacteria exist in culture-negative pediatric middle-ear effusions and that experimental infection with Haemophilus influenzae in the chinchilla model of otitis media results in the formation of adherent mucosal biofilms suggest that chronic OM may result from a mucosal biofilm infection.Objective To test the hypothesis that chronic OM in humans is biofilm-related.Design, Setting, and Patients Middle-ear mucosa (MEM) biopsy specimens were obtained from 26 children ( mean age, 2.5 [ range, 0.5-14] years) undergoing tympanostomy tube placement for treatment of otitis media with effusion (OME) and recurrent OM and were analyzed using microbiological culture, polymerase chain reaction (PCR)based diagnostics, direct microscopic examination, fluorescence in situ hybridization, and immunostaining. Uninfected ( control) MEM specimens were obtained from 3 children and 5 adults undergoing cochlear implantation. Patients were enrolled between February 2004 and April 2005 from a single US tertiary referral otolaryngology practice.Main Outcome Measures Confocal laser scanning microscopic (CLSM) images were obtained from MEM biopsy specimens and were evaluated for biofilm morphology using generic stains and species-specific probes for H influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis. Effusions, when present, were evaluated by PCR and culture for evidence of pathogen-specific nucleic acid sequences and bacterial growth, respectively.Results Of the 26 children undergoing tympanostomy tube placement, 13 (50%) had OME, 20 (77%) had recurrent OM, and 7 (27%) had both diagnoses; 27 of 52 (52%) of the ears had effusions, 24 of 24 effusions were PCR-positive for at least 1 OM pathogen, and 6 (22%) of 27 effusions were culture-positive for any pathogen. Mucosal biofilms were visualized by CLSM on 46 (92%) of 50 MEM specimens from children with OME and recurrent OM using generic and pathogen-specific probes. Biofilms were not observed on 8 control MEM specimens obtained from the patients undergoing cochlear implantation.Conclusion Direct detection of biofilms on MEM biopsy specimens from children with OME and recurrent OM supports the hypothesis that these chronic middle-ear disorders are biofilm-related.