MOLECULAR ANALYSIS OF BACTERIAL PATHOGENS IN OTITIS-MEDIA WITH EFFUSION
MOLECULAR ANALYSIS OF BACTERIAL PATHOGENS IN OTITIS-MEDIA WITH EFFUSION
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DOI：
10.1001/jama.273.20.1598
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发表时间：
1995-05-24
影响因子：
120.7
通讯作者：
EHRLICH, GD
中科院分区：
文献类型：
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作者：
POST, JC;PRESTON, RA;EHRLICH, GD
目的：确定聚合酶链式反应(PCR)能否检测经标准培养方法无菌的儿童中耳积液中的细菌DNA。设计-诊断方法的单中心、盲法比较研究。设计了卡他莫拉氏菌、流感嗜血杆菌和肺炎链球菌的聚合酶链式反应检测系统，并使用从培养细菌中获得的一组DNA进行了验证。收集慢性中耳积液标本，并通过培养和PCR设置进行对比分析。-三级保健儿科医院。患者。-从匹兹堡儿童医院的儿科门诊患者收集总共97例中耳积液(Pal在鼓膜切开和置管期间用于慢性中耳炎渗出性中耳炎(持续时间和>3个月)。结果：在97例渗出性中耳炎标本中，28例(28.9%)卡特氏支原体、流感嗜血杆菌和S肺炎支原体培养均为阳性(28.9%)，另有47例(48%)标本为卡他支原体、流感嗜血杆菌和S肺炎支原体培养阳性/培养阴性。因此，在97份检测样本中，75份(77.3%)对三种测试微生物中的一种或多种呈阳性。根据稀释实验，在平均培养阴性的耳朵中存在的细菌基因组等价物的最小数量估计大于10(4)。结论：基于PCR的检测系统可以检测到相当大比例的无培养中耳积液中细菌DNA的存在。虽然这一发现并不能证明活跃的细菌感染过程，但耳朵中存在的大量细菌基因组等价物表明存在活跃的感染过程。
Objective.-To determine if the polymerase chain reaction (PCR) can detect bacterial DNA in pediatric middle ear effusions that are sterile by standard cultural methods.Design.-Single-center, blinded, comparative study of diagnostic assays. The PCR-based detection systems for Moraxella catarrhalis, Haemophilus influenzae, and Streptococcus pneumoniae were designed and validated using a battery of DNAs obtained from cultured bacteria. Chronic middle ear effusion specimens were collected and comparatively analyzed by culture and the PCR.Setting.-Tertiary care pediatric hospital.Patients.-A total of 97 middle ear effusions were collected from pediatric outpatients at Children's Hospital of Pittsburgh (Pal during myringotomy and tube placement for chronic otitis media with effusion (duration >3 months). All patients had failed multiple courses of antimicrobial therapy and were diagnosed by a combination of validated otoscopy and tympanograms.Main Outcome Measure.-Differences in the percentage of positive test results between PCR-based assays and culture for M catarrhalis, H influenzae, and S pneumoniae.Results.-Of the 97 specimens of otitis media with effusion, 28 (28.9%) tested positive by both culture and PCR for M catarrhalis, H influenzae, or S pneumoniae, An additional 47 specimens (48%) were PCR positive/culture negative for these three bacterial species. Thus, 75 (77.3%) of the 97 specimens tested PCR positive for one or more of the three test organisms. The minimum number of bacterial genomic equivalents present in the average culture-negative ear was estimated to be greater than 10(4) based on dilutional experiments.Conclusions.-The PCR-based assay systems can detect the presence of bacterial DNA in a significant percentage of culturally sterile middle ear effusions. While this finding is not proof of an active bacterial infectious process, the large number of bacterial genomic equivalents present in the ears is suggestive of an active process.