Improved Detection of Biofilm-formative Bacteria by Vortexing and Sonication: A Pilot Study
Improved Detection of Biofilm-formative Bacteria by Vortexing and Sonication: A Pilot Study
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DOI：
10.1007/s11999-008-0609-5
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发表时间：
2009-05-01
影响因子：
4.2
通讯作者：
Bauer, Thomas W.
中科院分区：
文献类型：
--
作者：
Kobayashi, Hideo;Oethinger, Margret;Bauer, Thomas W.
骨科感染中常见的细菌（如葡萄球菌）形成生物膜并粘附在骨植入物和骨水泥上。已经报道了破坏生物膜和增强细菌检测的各种方法。我们将描述涡旋和超声处理的有效性，以提高传统的定量细菌培养和实时定量PCR检测生物膜形成细菌从聚甲基丙烯酸甲酯。我们使用单一的生物膜形成金黄色葡萄球菌菌株和20聚甲基丙烯酸甲酯试样作为体外生物膜模型;四个试样用于两个对照组或三个实验超声处理时间（1，5和30分钟）。在不进行超声处理的情况下涡旋骨水泥在相当大的程度上增加了粘附细菌的产量。无论超声处理的持续时间如何，涡旋和超声处理的组合都进一步提高了产率。定量常规培养与定量PCR测定相关。结合涡旋和超声处理破坏细菌生物膜，然后进行定量PCR和/或培养，似乎是检测粘附在骨水泥上的细菌的敏感方法。
Bacteria such as staphylococci commonly encountered in orthopaedic infections form biofilms and adhere to bone implants and cements. Various methods to disrupt the biofilm and enhance bacterial detection have been reported. We will describe the effectiveness of vortexing and sonication to improve the detection of biofilm-formative bacteria from polymethylmethacrylate by conventional quantitative bacterial culture and real-time quantitative PCR. We used a single biofilm-formative Staphylococcus aureus strain and 20 polymethylmethacrylate coupons as an in vitro biofilm model; four coupons were used for each of two control groups or three experimental sonication times (1, 5, and 30 minutes). Vortexing the cement without sonication increased the yield of adherent bacteria to a considerable extent. The combination of vortexing and sonication further enhanced the yield regardless of the duration of sonication. Quantitative conventional cultures correlated with quantitative PCR assay. The combination of vortexing and sonication to disrupt the bacterial biofilm followed by quantitative PCR and/or culture seems to be a sensitive method for detecting bacteria adherent to bone cement.