Skip to main content

Advertisement

SpringerLink
Log in

Molecular Identification of Bacteria from Aseptically Loose Implants

  • Original Article
  • Published:
Clinical Orthopaedics and Related Research

Abstract

Polymerase chain reaction (PCR) assays have been used to detect bacteria adherent to failed orthopaedic implants, but some PCR assays have had problems with probable false-positive results. We used a combination of a Staphylococcus species-specific PCR and a universal PCR followed by DNA sequencing to identify bacteria on implants retrieved from 52 patients (92 implants) at revision arthroplasty. We addressed two questions in this study: (1) Is this method able to show the existence of bacterial DNA on presumed aseptic loosed implants?; and (2) What proportion of presumed aseptic or culture-negative implants was positive for bacterial DNA by PCR? Fourteen implants (15%) were believed infected, whereas 74 implants (85%) were believed aseptic. Each implant was sonicated and the resulting solution was submitted for dual real-time PCR assay and culture. All implants believed aseptically loose were culture-negative, but nine of the 74 (12%) had bacterial DNA by PCR; two (2.7%) were PCR-positive and also showed histologic findings suggestive of infection. Uniquely developed PCR and bacterial sequencing assays showed bacterial DNA on 12% of implants removed for presumed aseptic loosening. Additional studies are needed to determine the clinical importance of bacterial DNA detected by PCR but not by conventional culture.

Level of Evidence: Level III, diagnostic study. See the Guidelines for Authors for a complete description of levels of evidence.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Arciola CR, Campoccia D, Montanaro L. Detection of biofilm-forming strains of Staphylococcus epidermidis and S. aureus. Expert Rev Mol Diagn. 2002;2:478–484.

    Article  PubMed  CAS  Google Scholar 

  2. Bauer TW, Parvizi J, Kobayashi N, Krebs V. Diagnosis of periprosthetic infection. J Bone Joint Surg Am. 2006;88:869–882.

    Article  PubMed  Google Scholar 

  3. Borst A, Box AT, Fluit AC. False-positive results and contamination in nucleic acid amplification assays: suggestions for a prevent and destroy strategy. Eur J Clin Microbiol Infect Dis. 2004;23:289–299.

    Article  PubMed  CAS  Google Scholar 

  4. Clarke MT, Roberts CP, Lee PT, Gray J, Keene GS, Rushton N. Polymerase chain reaction can detect bacterial DNA in aseptically loose total hip arthroplasties. Clin Orthop Relat Res. 2004;427:132–137.

    Article  PubMed  Google Scholar 

  5. Clohisy JC, Calvert G, Tull F, McDonald D, Maloney WJ. Reasons for revision hip surgery: a retrospective review. Clin Orthop Relat Res. 2004;429:188–192.

    Article  PubMed  Google Scholar 

  6. Corless CE, Guiver M, Borrow R, Edwards-Jones V, Kaczmarski EB, Fox AJ. Contamination and sensitivity issues with a real-time universal 16S rRNA PCR. J Clin Microbiol. 2000;38:1747–1752.

    PubMed  CAS  Google Scholar 

  7. Espehaug B, Engesaeter LB, Vollset SE, Havelin LI, Langeland N. Antibiotic prophylaxis in total hip arthroplasty: review of 10,905 primary cemented total hip replacements reported to the Norwegian arthroplasty register, 1987 to 1995. J Bone Joint Surg Br. 1997;79:590–595.

    Article  PubMed  CAS  Google Scholar 

  8. Grahn N, Olofsson M, Ellnebo-Svedlund K, Monstein HJ, Jonasson J. Identification of mixed bacterial DNA contamination in broad-range PCR amplification of 16S rDNA V1 and V3 variable regions by pyrosequencing of cloned amplicons. FEMS Microbiol Lett. 2003;219:87–91.

    Article  PubMed  CAS  Google Scholar 

  9. Hirakawa K, Jacobs JJ, Urban R, Saito T. Mechanisms of failure of total hip replacements: lessons learned from retrieval studies. Clin Orthop Relat Res. 2004;420:10–17.

    Article  PubMed  Google Scholar 

  10. Ince A, Rupp J, Frommelt L, Katzer A, Gille J, Lohr JF. Is “aseptic” loosening of the prosthetic cup after total hip replacement due to nonculturable bacterial pathogens in patients with low-grade infection? Clin Infect Dis. 2004;39:1599–1603.

    Article  PubMed  CAS  Google Scholar 

  11. Josefsson G, Lindberg L, Wiklander B. Systemic antibiotics and gentamicin-containing bone cement in the prophylaxis of postoperative infections in total hip arthroplasty. Clin Orthop Relat Res. 1981;159:194–200.

    PubMed  Google Scholar 

  12. Kobayashi N, Bauer TW, Sakai H, Togawa D, Lieberman IH, Fujishiro T, Procop GW. The use of newly developed real-time PCR for the rapid identification of bacteria in culture-negative osteomyelitis: a case report. Joint Bone Spine. 2006;73:745–747.

    Article  PubMed  Google Scholar 

  13. Kobayashi N, Bauer TW, Togawa D, Lieberman IH, Sakai H, Fujishiro T, Tuohy MJ, Procop GW. A molecular Gram stain using broad range PCR and pyrosequencing technology: a potentially useful tool for diagnosing orthopaedic infections. Diagn Mol Pathol. 2005;14:83–89.

    Article  PubMed  CAS  Google Scholar 

  14. Kobayashi N, Bauer TW, Tuohy MJ, Fujishiro T, Procop GW. Brief ultrasonication improves detection of biofilm-formative bacteria around a metal implant. Clin Orthop Relat Res. 2007;457:210–213.

    PubMed  Google Scholar 

  15. Kobayashi N, Bauer TW, Tuohy MJ, Lieberman IH, Krebs V, Togawa D, Procop GW. The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections. J Orthop Res. 2006;24:1641–1649.

    Article  PubMed  CAS  Google Scholar 

  16. Malchau H, Herberts P, Eisler T, Garellick G, Soderman P. The Swedish Total Hip Replacement Register. J Bone Joint Surg Am. 2002;84(suppl 2):2–20.

    PubMed  Google Scholar 

  17. Mariani BD, Martin DS, Levine MJ, Booth RE Jr, Tuan RS. The Coventry Award. Polymerase chain reaction detection of bacterial infection in total knee arthroplasty. Clin Orthop Relat Res. 1996;331:11–22.

    Article  PubMed  Google Scholar 

  18. Mirra JM, Amstutz HC, Matos M, Gold R. The pathology of the joint tissues and its clinical relevance in prosthesis failure. Clin Orthop Relat Res. 1976;117:221–240.

    PubMed  Google Scholar 

  19. Nelson CL, McLaren AC, McLaren SG, Johnson JW, Smeltzer MS. Is aseptic loosening truly aseptic? Clin Orthop Relat Res. 2005;437:25–30.

    Article  PubMed  Google Scholar 

  20. Neut D, van Horn JR, van Kooten TG, van der Mei HC, Busscher HJ. Detection of biomaterial-associated infections in orthopaedic joint implants. Clin Orthop Relat Res. 2003;413:261–268.

    Article  PubMed  Google Scholar 

  21. Nguyen LL, Nelson CL, Saccente M, Smeltzer MS, Wassell DL, McLaren SG. Detecting bacterial colonization of implanted orthopaedic devices by ultrasonication. Clin Orthop Relat Res. 2002;403:29–37.

    Article  PubMed  Google Scholar 

  22. Sakai H, Procop GW, Kobayashi N, Togawa D, Wilson DA, Borden L, Krebs V, Bauer TW. Simultaneous detection of Staphylococcus aureus and coagulase-negative staphylococci in positive blood cultures by real-time PCR with two fluorescence resonance energy transfer probe sets. J Clin Microbiol. 2004;42:5739–5744.

    Article  PubMed  CAS  Google Scholar 

  23. Schmalzried TP, Kwong LM, Jasty M, Sedlacek RC, Haire TC, O’Connor DO, Bragdon CR, Kabo JM, Malcolm AJ, Harris WH. The mechanism of loosening of cemented acetabular components in total hip arthroplasty: analysis of specimens retrieved at autopsy. Clin Orthop Relat Res. 1992;274:60–78.

    PubMed  Google Scholar 

  24. Stoodley P, Kathju S, Hu FZ, Erdos G, Levenson JE, Mehta N, Dice B, Johnson S, Hall-Stoodley L, Nistico L, Sotereanos N, Sewecke J, Post JC, Ehrlich GD. Molecular and imaging techniques for bacterial biofilms in joint arthroplasty infections. Clin Orthop Relat Res. 2005;437:31–40.

    Article  PubMed  Google Scholar 

  25. Trampuz A, Osmon DR, Hanssen AD, Steckelberg JM, Patel R. Molecular and antibiofilm approaches to prosthetic joint infection. Clin Orthop Relat Res. 2003;414:69–88.

    Article  PubMed  Google Scholar 

  26. Tunney MM, Patrick S, Curran MD, Ramage G, Anderson N, Davis RI, Gorman SP, Nixon JR. Detection of prosthetic joint biofilm infection using immunological and molecular techniques. Methods Enzymol. 1999;310:566–576.

    Article  PubMed  CAS  Google Scholar 

  27. Tunney MM, Patrick S, Curran MD, Ramage G, Hanna D, Nixon JR, Gorman SP, Davis RI, Anderson N. Detection of prosthetic hip infection at revision arthroplasty by immunofluorescence microscopy and PCR amplification of the bacterial 16 rRNA gene. J Clin Microbiol. 1999;37:3281–3290.

    PubMed  CAS  Google Scholar 

  28. Tunney MM, Patrick S, Gorman SP, Nixon JR, Anderson N, Davis RI, Hanna D, Ramage G. Improved detection of infection in hip replacements: a currently underestimated problem. J Bone Joint Surg Br. 1998;80:568–572.

    Article  PubMed  CAS  Google Scholar 

  29. van de Belt H, Neut D, Schenk W, van Horn JR, van der Mei HC, Busscher HJ. Infection of orthopedic implants and the use of antibiotic-loaded bone cements: a review. Acta Orthop Scand. 2001;72:557–571.

    Article  PubMed  Google Scholar 

  30. Yang S, Rothman RE. PCR-based diagnostics for infectious diseases: uses, limitations, and future applications in acute-care settings. Lancet Infect Dis. 2004;4:337–348.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomic Pathology, L25, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

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

  2. Department of Orthopaedic Surgery, The Cleveland Clinic Foundation, Cleveland, OH, USA

    Naomi Kobayashi MD, PhD, Viktor Krebs MD, Hideo Kobayashi MD, PhD & Thomas W. Bauer MD, PhD

  3. Department of Orthopaedic Surgery, Yokohama City University School of Medicine, Yokohama, Japan

    Naomi Kobayashi MD, PhD & Hideo Kobayashi MD, PhD

  4. Department of Clinical Microbiology, The Cleveland Clinic Foundation, Cleveland, OH, USA

    Gary W. Procop MD

Authors
  1. Naomi Kobayashi MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gary W. Procop MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Viktor Krebs MD
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hideo Kobayashi MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Thomas W. Bauer MD, PhD
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Thomas W. Bauer MD, PhD.

Additional information

One or more of the authors (TW) has received funding from Stryker Orthopedics (Mahwah, NJ) and Nippon Stryker KK (Tokyo, Japan).

About this article

Cite this article

Kobayashi, N., Procop, G.W., Krebs, V. et al. Molecular Identification of Bacteria from Aseptically Loose Implants. Clin Orthop Relat Res 466, 1716–1725 (2008). https://doi.org/10.1007/s11999-008-0263-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11999-008-0263-y

Keywords

Search

Navigation