Can We Trust Intraoperative Culture Results in Nonunions?
Can We Trust Intraoperative Culture Results in Nonunions?
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DOI：
10.1097/bot.0000000000000043
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发表时间：
2014-07-01
影响因子：
2.3
通讯作者：
Costerton, John W.
中科院分区：
文献类型：
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作者：
Palmer, Michael P.;Altman, Daniel T.;Costerton, John W.
Objectives：为了确定骨不连中细菌生物膜的存在，比较分子技术（多重聚合酶链反应和质谱，荧光原位杂交）与常规术中culture.Methods：34例骨不连患者计划进行手术，并参加了这项正在进行的前瞻性研究。从取出的植入物、周围组织膜和局部软组织采集术中标本，然后进行标准培养分析、Ibis第二代分子诊断（Ibis Biosystems）和细菌16S rRNA荧光原位杂交（FISH）。共聚焦显微镜被用来可视化的组织标本与FISH探针，这是根据选择的Ibis analysis.Results：34例患者的遭遇进行了分析。8例术中培养阳性，诊断为感染性骨不连。Ibis确认了所有8个样本中存在细菌。Ibis在34次接触中总共发现了30次细菌，这些数据得到了FISH的证实。30份Ibis阳性样本中有22份培养阴性。所有分析方法均显示4份样本为阴性。没有样品是阳性的文化，但阴性的分子technology.Conclusions：我们的初步数据表明，分子诊断是更敏感的识别细菌比文化的情况下，骨不连。这可能是因为培养物无法检测生物膜和先前暴露于抗生素治疗的细菌。
Objectives:To identify the presence of bacterial biofilms in nonunions comparing molecular techniques (multiplex polymerase chain reaction and mass spectrometry, fluorescent in situ hybridization) with routine intraoperative cultures.Methods:Thirty-four patients with nonunions were scheduled for surgery and enrolled in this ongoing prospective study. Intraoperative specimens were collected from removed implants, surrounding tissue membrane, and local soft tissue followed by standard culture analysis, Ibis's second generation molecular diagnostics (Ibis Biosystems), and bacterial 16S rRNA-based fluorescence in situ hybridization (FISH). Confocal microscopy was used to visualize the tissue specimens reacted with the FISH probes, which were chosen based on the Ibis analysis.Results:Thirty-four patient encounters were analyzed. Eight were diagnosed as infected nonunions by positive intraoperative culture results. Ibis confirmed the presence of bacteria in all 8 samples. Ibis identified bacteria in a total of 30 of 34 encounters, and these data were confirmed by FISH. Twenty-two of 30 Ibis-positive samples were culture-negative. Four samples were negative by all methods of analysis. No samples were positive by culture, but negative by molecular techniques.Conclusions:Our preliminary data indicate that molecular diagnostics are more sensitive for identifying bacteria than cultures in cases of bony nonunion. This is likely because of the inability of cultures to detect biofilms and bacteria previously exposed to antibiotic therapy.