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Improved Detection of Biofilm-formative Bacteria by Vortexing and Sonication: A Pilot Study

Authors
  • Kobayashi, Hideo1, 2
  • Oethinger, Margret1
  • Tuohy, Marion J.1
  • Procop, Gary W.1
  • Bauer, Thomas W.1, 2, 3
  • 1 The Cleveland Clinic, Institute of Pathology and Laboratory Medicine, Cleveland, OH, USA , Cleveland (United States)
  • 2 The Cleveland Clinic, Orthopaedic and Rheumatologic Institute, Cleveland, OH, USA , Cleveland (United States)
  • 3 L-25, The Cleveland Clinic Foundation, Departments of Anatomic Pathology, Orthopaedic Surgery, and the Spine Institute, 9500 Euclid Avenue, Cleveland, OH, 44195, USA , Cleveland (United States)
Type
Published Article
Journal
Clinical Orthopaedics and Related Research
Publisher
Ovid Technologies (Wolters Kluwer) - Lippincott Williams & Wilkins
Publication Date
Nov 07, 2008
Volume
467
Issue
5
Pages
1360–1364
Identifiers
DOI: 10.1007/s11999-008-0609-5
Source
Springer Nature
Keywords
License
Yellow

Abstract

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.

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