Microscopic Detection of Viable Staphylococcus epidermidis in Peri-Implant Tissue in Experimental Biomaterial-Associated Infection, Identified by Bromodeoxyuridine Incorporation
Microscopic Detection of Viable Staphylococcus epidermidis in Peri-Implant Tissue in Experimental Biomaterial-Associated Infection, Identified by Bromodeoxyuridine Incorporation
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DOI：
10.1128/iai.00849-09
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发表时间：
2010-03-01
影响因子：
3.1
通讯作者：
Zaat, S. A. J.
中科院分区：
文献类型：
--
作者：
Broekhuizen, C. A. N.;Sta, M.;Zaat, S. A. J.
生物医学器械感染的特征在于生物膜形成和病原体在周围组织中的定殖。为了研究在感染器械周围组织中显微镜下检测到的细菌是否存活，我们使用了溴脱氧尿苷（BrdU），这是一种掺入细菌DNA中的核苷酸类似物，可以用抗体检测到。受感染的人体组织是从植入血管内器械的患者死后获得的，小鼠活检标本是从实验性生物材料感染的小鼠获得的。体外实验表明，表皮葡萄球菌纳入BrdU，从细菌与抗BrdU抗体染色判断。在将细菌与BrdU孵育并随后用抗BrdU抗体对显微镜切片进行染色后，可以在死亡患者的血管内装置周围的组织中清楚地观察到细菌。利用这种染色技术，可以在用低剂量S.在攻击后14 - 21天用地塞米松处理以抑制免疫。这证实并扩展了我们先前的发现，即导管周围组织是生物材料相关感染中细菌的储存库。感染的发病机制和时间-空间分布的活的，分裂的细菌，现在可以在显微镜水平上进行研究，通过免疫标记与BrdU和BrdU抗体。
Infection of biomedical devices is characterized by biofilm formation and colonization of surrounding tissue by the causative pathogens. To investigate whether bacteria detected microscopically in tissue surrounding infected devices were viable, we used bromodeoxyuridine (BrdU), a nucleotide analogue that is incorporated into bacterial DNA and can be detected with antibodies. Infected human tissue was obtained postmortem from patients with intravascular devices, and mouse biopsy specimens were obtained from mice with experimental biomaterial infection. In vitro experiments showed that Staphylococcus epidermidis incorporated BrdU, as judged from staining of the bacteria with anti-BrdU antibodies. After incubation of bacteria with BrdU and subsequent staining of microscopic sections with anti-BrdU antibodies, bacteria could be clearly visualized in the tissue surrounding intravascular devices of deceased patients. With this staining technique, relapse of infection could be visualized in mice challenged with a low dose of S. epidermidis and treated with dexamethasone between 14 and 21 days after challenge to suppress immunity. This confirms and extends our previous findings that pericatheter tissue is a reservoir for bacteria in biomaterial-associated infection. The pathogenesis of the infection and temporo-spatial distribution of viable, dividing bacteria can now be studied at the microscopic level by immunolabeling with BrdU and BrdU antibodies.