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

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Clinical Orthopaedics and Related Research

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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Acknowledgments

We thank Dr. Gerri S. Hall for assistance in preparing the manuscript. We also are grateful to Benjamin Nutter for advice on the statistical analysis.

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Authors and Affiliations

  1. Institute of Pathology and Laboratory Medicine, The Cleveland Clinic, Cleveland, OH, USA

    Hideo Kobayashi MD, PhD, Margret Oethinger MD, PhD, Marion J. Tuohy MT (ASCP), Gary W. Procop MS, MD & Thomas W. Bauer MD, PhD

  2. Orthopaedic and Rheumatologic Institute, The Cleveland Clinic, Cleveland, OH, USA

    Hideo Kobayashi MD, PhD & Thomas W. Bauer MD, PhD

  3. Departments of Anatomic Pathology, Orthopaedic Surgery, and the Spine Institute, L-25, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

    Thomas W. Bauer MD, PhD

Authors
  1. Hideo Kobayashi MD, PhD
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  2. Margret Oethinger MD, PhD
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  3. Marion J. Tuohy MT (ASCP)
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  4. Gary W. Procop MS, MD
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  5. Thomas W. Bauer MD, PhD
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Corresponding author

Correspondence to Thomas W. Bauer MD, PhD.

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Each author certifies that he or she has no commercial associations (eg, consultancies, stock ownership, equity interest, patent/licensing arrangements, etc) that might pose a conflict of interest in connection with the submitted article.

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Kobayashi, H., Oethinger, M., Tuohy, M.J. et al. Improved Detection of Biofilm-formative Bacteria by Vortexing and Sonication: A Pilot Study. Clin Orthop Relat Res 467, 1360–1364 (2009). https://doi.org/10.1007/s11999-008-0609-5

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  • DOI: https://doi.org/10.1007/s11999-008-0609-5

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