Skip to main content

Advertisement

SpringerLink
Log in

Molecular detection of transcriptionally active bacteria from failed prosthetic hip joints removed during revision arthroplasty

  • Article
  • Published:
European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

The purpose of this study was to use microbiological culture and bacterial 16S rRNA gene sequencing methods to detect transcriptionally active bacteria present on the surface of failed prosthetic hip joints removed during revision arthroplasty. Five failed prosthetic hip joints were sonicated to dislodge adherent bacteria and subjected to microbiological culture. Bacterial RNA was extracted from each sonicate, cDNA prepared by reverse transcription and the 16S rRNA gene amplified using universal primers. Polymerase chain reaction (PCR) products were cloned, assigned to distinct groups by restriction fragment length polymorphism (RFLP) analysis and one representative clone from each group was sequenced. Bacteria were identified by comparison of the obtained 16S rRNA gene sequences with those deposited in public access sequence databases. All five specimens were positive for the presence of bacteria by both culture and PCR. Culture methods identified species from eight genera. Molecular detection of transcriptionally active bacteria identified a wider range of species. A total of 42 phylotypes were identified, of which Lysobacter gummosus was the most abundant (31.6%). Thirty-four clones (14.5%) represented uncultivable phylotypes. No potentially novel species were identified. It is concluded that a diverse range of transcriptionally active bacterial species are present within biofilms on the surface of failed prosthetic hip joints.

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

Similar content being viewed by others

References

  1. Patel R, Osmon DR, Hanssen AD (2005) The diagnosis of prosthetic joint infection: current techniques and emerging technologies. Clin Orthop Relat Res 437:55–58

    Article  PubMed  Google Scholar 

  2. Toms AD, Davidson D, Masri BA, Duncan CP (2006) The management of peri-prosthetic infection in total joint arthroplasty. J Bone Joint Surg Br 88:149–155

    CAS  PubMed  Google Scholar 

  3. NHS Direct Health Encyclopaedia. Hip replacement. Available online at: http://www.nhsdirect.nhs.uk/articles/article.aspx?articleId=522

  4. Lentino JR (2003) Prosthetic joint infections: bane of orthopedists, challenge for infectious disease specialists. Clin Infect Dis 36:1157–1161

    Article  PubMed  Google Scholar 

  5. Lachiewicz PF, Rogers GD, Thomason HC (1996) Aspiration of the hip joint before revision total hip arthroplasty. Clinical and laboratory factors influencing attainment of a positive culture. J Bone Joint Surg Am 78:749–754

    CAS  PubMed  Google Scholar 

  6. Thomas JG, Ramage G, Lopez-Ribot JL (2004) Biofilms and implant infections. In: Ghannoum M, O’Toole G (eds) Microbial biofilms. American Society of Microbiology Press, Washington, pp 269–293

    Google Scholar 

  7. Atkins BL, Athanasou N, Deeks JJ, Crook DWM, Simpson H, Peto TEA, McLardy-Smith P, Berendt AR (1998) Prospective evaluation of criteria for microbiological diagnosis of prosthetic-joint infection at revision arthroplasty. The OSIRIS Collaborative Study Group. J Clin Microbiol 36:2932–2939

    CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  9. Dempsey KE, Riggio MP, Lennon A, Hannah VE, Ramage G, Allan D, Bagg J (2007) Identification of bacteria on the surface of clinically infected and non-infected prosthetic hip joints removed during revision arthroplasties by 16S rRNA gene sequencing and by microbiological culture. Arthritis Res Ther 9:R46

    Article  PubMed  Google Scholar 

  10. Trampuz A, Piper KE, Jacobson MJ, Hanssen AD, Unni KK, Osmon DR, Mandrekar JN, Cockerill FR, Steckelberg JM, Greenleaf JF, Patel R (2007) Sonication of removed hip and knee prostheses for diagnosis of infection. N Engl J Med 357:654–663

    Article  CAS  PubMed  Google Scholar 

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

    Article  PubMed  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  13. Mariani BD, Tuan RS (1998) Advances in the diagnosis of infection in prosthetic joint implants. Mol Med Today 4:207–213

    Article  CAS  PubMed  Google Scholar 

  14. Levine MJ, Mariani BA, Tuan RS, Booth RE Jr (1995) Molecular genetic diagnosis of infected total joint arthroplasty. J Arthroplasty 10:93–94

    Article  CAS  PubMed  Google Scholar 

  15. Cox CJ, Kempsell KE, Hill Gaston JS (2003) Investigation of infectious agents associated with arthritis by reverse transcription PCR of bacterial rRNA. Arthritis Res Ther 5:R1–R8

    Article  CAS  PubMed  Google Scholar 

  16. Kempsell KE, Cox CJ, Hurle M, Wong A, Wilkie S, Zanders ED, Hill Gaston JS, Crowe JS (2000) Reverse transcriptase-PCR analysis of bacterial rRNA for detection and characterization of bacterial species in arthritis synovial tissue. Infect Immun 68:6012–6026

    Article  CAS  PubMed  Google Scholar 

  17. Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. John Wiley and Sons, New York, pp 115–175

    Google Scholar 

  18. Riggio MP, Lennon A, Wray D (2000) Detection of Helicobacter pylori DNA in recurrent aphthous stomatitis tissue by PCR. J Oral Pathol Med 29:507–513

    Article  CAS  PubMed  Google Scholar 

  19. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402

    Article  CAS  PubMed  Google Scholar 

  20. Shimizu A, Akiyama M, Ishijima Y, Hama K, Kunimaru T, Naganuma T (2006) Molecular characterization of microbial communities in fault-bordered aquifers in the Miocene formation of northernmost Japan. Geobiol 4:203–213

    Article  CAS  Google Scholar 

  21. Bartzokas CA, Johnson R, Jane M, Martin MV, Pearce PK, Saw Y (1994) Relation between mouth and haematogenous infection in total joint replacements. Br Med J 309:506–508

    CAS  Google Scholar 

  22. Stoll T, Stucki G, Brühlmann P, Vogt M, Gschwend N, Michel BA (1996) Infection of a total knee joint prosthesis by Peptostreptococcus micros and Propionibacterium acnes in an elderly RA patient: implant salvage with longterm antibiotics and needle aspiration/irrigation. Clin Rheumatol 15:399–402

    Article  CAS  PubMed  Google Scholar 

  23. Waldman BJ, Mont MA, Hungerford DS (1997) Total knee arthroplasty infections associated with dental procedures. Clin Orthop Relat Res 343:164–172

    Article  PubMed  Google Scholar 

  24. LaPorte DM, Waldman BJ, Mont MA, Hungerford DS (1999) Infections associated with dental procedures in total hip arthroplasty. J Bone Joint Surg Br 81:56–59

    Article  CAS  PubMed  Google Scholar 

  25. Tronstad L, Sunde PT (2003) The evolving new understanding of endodontic infections. Endod Top 6:57–77

    Article  Google Scholar 

  26. Lockhart PB, Loven B, Brennan MT, Fox PC (2007) The evidence base for the efficacy of antibiotic prophylaxis in dental practice. J Am Dent Assoc 138:458–474

    PubMed  Google Scholar 

  27. Fenollar F, Roux V, Stein A, Drancourt M, Raoult D (2006) Analysis of 525 samples to determine the usefulness of PCR amplification and sequencing of the 16S rRNA gene for diagnosis of bone and joint infections. J Clin Microbiol 44:1018–1028

    Article  CAS  PubMed  Google Scholar 

  28. Zimmerli W, Trampuz A, Ochsner PE (2004) Prosthetic-joint infections. N Engl J Med 351:1645–1654

    Article  CAS  PubMed  Google Scholar 

  29. Panousis K, Grigoris P, Butcher I, Rana B, Reilly JH, Hamblen DL (2005) Poor predictive value of broad-range PCR for the detection of arthroplasty infection in 92 cases. Acta Orthop 76:341–346

    PubMed  Google Scholar 

  30. An SY, Asahara M, Goto K, Kasai H, Yokota A (2007) Virgibacillus halophilus sp. nov., spore-forming bacteria isolated from soil in Japan. Int J Syst Evol Microbiol 57:1607–1611

    Article  PubMed  Google Scholar 

  31. Graur D, Pupko T (2001) The Permian bacterium that isn’t. Mol Biol Evol 18:1143–1146

    CAS  PubMed  Google Scholar 

  32. D’Amico M, Mangano S, Spinelli M, Sala E, Vigano EF, Grilli R, Fraticelli M, Grillo C, Limido A (2005) Epidemic of infections caused by ‘aquatic’ bacteria in patients undergoing hemodialysis via central venous catheters. G Ital Nefrol 22:508–513

    PubMed  Google Scholar 

  33. Chi C-Y, Fung C-P, Wong W-W, Liu C-Y (2004) Brevundimonas bacteremia: two case reports and literature review. Scand J Infect Dis 36:59–61

    Article  PubMed  Google Scholar 

  34. Sofer Y, Zmira S, Amir J (2007) Brevundimonas vesicularis septic arthritis in an immunocompetent child. Eur J Pediatr 166:77–78

    Article  PubMed  Google Scholar 

  35. Marques da Silva R, Caugant DA, Eribe ERK, Aas JA, Lingaas PS, Geiran O, Tronstad L, Olsen I (2006) Bacterial diversity in aortic aneurysms determined by 16S ribosomal RNA gene analysis. J Vasc Surg 44:1055–1060

    Article  PubMed  Google Scholar 

  36. Paster BJ, Falkler WA Jr, Enwonwu CO, Idigbe EO, Savage KO, Levanos VA, Tamer MA, Ericson RL, Lau CN, Dewhirst FE (2002) Prevalent bacterial species and novel phylotypes in advanced noma lesions. J Clin Microbiol 40:2187–2191

    Article  CAS  PubMed  Google Scholar 

  37. Wildeboer-Veloo ACM, Harmsen HJM, Welling GW, Degener JE (2007) Development of 16S rRNA-based probes for the identification of Gram-positive anaerobic cocci isolated from human clinical specimens. Clin Microbiol Infect 13:985–992

    Article  CAS  PubMed  Google Scholar 

  38. Song Y, Liu C, Finegold SM (2007) Peptoniphilus gorbachii sp. nov., Peptoniphilus olsenii sp. nov., and Anaerococcus murdochii sp. nov. isolated from clinical specimens of human origin. J Clin Microbiol 45:1746–1752

    Article  CAS  PubMed  Google Scholar 

  39. Looney WJ (2005) Role of Stenotrophomonas maltophilia in hospital-acquired infection. Br J Biomed Sci 62:145–154

    CAS  PubMed  Google Scholar 

  40. Guilhabert MR, Kirkpatrick BC (2005) Identification of Xylella fastidiosa antivirulence genes: hemagglutinin adhesins contribute to X. fastidiosa biofilm maturation and colonization and attenuate virulence. Mol Plant Microbe Interact 18:856–868

    Article  CAS  PubMed  Google Scholar 

  41. Jacques M-A, Josi K, Darrasse A, Samson R (2005) Xanthomonas axonopodis pv. phaseoli var. fuscans is aggregated in stable biofilm population sizes in the phyllosphere of field-grown beans. Appl Environ Microbiol 71:2008–2015

    Article  CAS  PubMed  Google Scholar 

  42. Huang T-P, Somers EB, Wong ACL (2006) Differential biofilm formation and motility associated with lipopolysaccharide/exopolysaccharide-coupled biosynthetic genes in Stenotrophomonas maltophilia. J Bacteriol 188:3116–3120

    Article  CAS  PubMed  Google Scholar 

  43. Kämpfer P, Rosselló-Mora R, Hermansson M, Persson F, Huber B, Falsen E, Busse HJ (2007) Undibacterium pigrum gen. nov., sp. nov., isolated from drinking water. Int J Syst Evol Microbiol 57:1510–1515

    Article  PubMed  Google Scholar 

  44. Simões LC, Simões M, Vieira MJ (2007) Biofilm interactions between distinct bacterial genera isolated from drinking water. Appl Environ Microbiol 73:6192–6200

    Article  PubMed  Google Scholar 

  45. Saxena N, Misra R, Aggarwal A (2006) Is the enthesitis-related arthritis subtype of juvenile idiopathic arthritis a form of chronic reactive arthritis? Rheumatology 45:1129–1132

    Article  CAS  PubMed  Google Scholar 

  46. Abraham WR, Strömpl C, Meyer H, Lindholst S, Moore ER, Christ R, Vancanneyt M, Tindall BJ, Bennasar A, Smit J, Tesar M (1999) Phylogeny and polyphasic taxonomy of Caulobacter species. Proposal of Maricaulis gen. nov. with Maricaulis maris (Poindexter) comb. nov. as the type species, and emended description of the genera Brevundimonas and Caulobacter. Int J Syst Bacteriol 49:1053–1073

    Article  CAS  PubMed  Google Scholar 

  47. Smith JL, Fratamico PM, Gunther NW (2007) Extraintestinal pathogenic Escherichia coli. Foodborne Pathog Dis 4:134–163

    Article  CAS  PubMed  Google Scholar 

  48. Falagas ME, Karveli EA, Kelesidis I, Kelesidis T (2007) Community-acquired Acinetobacter infections. Eur J Microbiol Infect Dis 26:857–868

    Article  CAS  Google Scholar 

  49. Davies JC, Rubin BK (2007) Emerging and unusual gram-negative infections in cystic fibrosis. Semin Respir Crit Car Med 28:312–321

    Article  Google Scholar 

  50. Torres-Tortosa M, Arrizabalaga J, Villanueva JL, Gálvez J, Leyes M, Valencia ME, Flores J, Peña JM, Pérez-Cecilia E, Quereda C (2003) Prognosis and clinical evaluation of infection caused by Rhodococcus equi in HIV-infected patients: a multicenter study of 67 cases. Chest 123:1970–1976

    Article  PubMed  Google Scholar 

  51. Rozsypal H, Aster V, Stahiková M, Horová B (2007) Rhodococcus equi infection in subjects infected with human immunodeficiency virus (HIV). Cas Lek Cesk 146:163–167

    CAS  PubMed  Google Scholar 

  52. North NN, Dollhopf SL, Petrie L, Istok JD, Balkwill DL, Kostka JE (2004) Change in bacterial community structure during in situ biostimulation of subsurface sediment cocontaminated with uranium and nitrate. Appl Environ Microbiol 70:4911–4920

    Article  CAS  PubMed  Google Scholar 

  53. Huber JA, Johnson HP, Butterfield DA, Baross JA (2006) Microbial life in ridge flank crustal fluids. Environ Microbiol 8:88–99

    Article  CAS  PubMed  Google Scholar 

  54. Kaiser O, Pühler A, Selbitschka W (2001) Phylogenetic analysis of microbial diversity in the rhizoplane of oilseed rape (Brassica napus cv. Westar) employing cultivation-dependent and cultivation-independent approaches. Microb Ecol 42:136–149

    CAS  PubMed  Google Scholar 

  55. Rawls JF, Mahowald MA, Ley RE, Gordon JI (2006) Reciprocal gut microbiota transplants from zebrafish and mice to germ-free recipients reveal host habitat selection. Cell 127:423–433

    Article  CAS  PubMed  Google Scholar 

  56. Williams MM, Domingo JWS, Meckes MC, Kelty CA, Rochon HS (2004) Phylogenetic diversity of drinking water bacteria in a distribution system simulator. J Appl Microbiol 96:954–964

    Article  CAS  PubMed  Google Scholar 

  57. Flanagan JL, Brodie EL, Weng L, Lynch SV, Garcia O, Brown R, Hugenholtz P, DeSantis TZ, Andersen GL, Wiener-Kronish JP, Bristow J (2007) Loss of bacterial diversity during antibiotic treatment of intubated patients colonized with Pseudomonas aeruginosa. J Clin Microbiol 45:1954–1962

    Article  CAS  PubMed  Google Scholar 

  58. Oliver JD (2005) The viable but nonculturable state in bacteria. J Microbiol 43 Spec No:93–100

    Google Scholar 

  59. Giesler LJ, Yuen GY (1998) Evaluation of Stenotrophomonas maltophilia strain C3 for biocontrol of brown patch disease. Crop Prot 17:509–513

    Article  Google Scholar 

  60. Senol E (2004) Stenotrophomonas maltophilia: the significance and role as a nosocomial pathogen. J Hosp Infect 57:1–7

    Article  CAS  PubMed  Google Scholar 

  61. Denton M, Kerr KG (1998) Microbiological and clinical aspects of infection associated with Stenotrophomonas maltophilia. Clin Microbiol Rev 11:57–80

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Dr. Dominic Meek for the provision of the prosthetic hip joint samples and Dr. Grace Sweeney for conducting bacteriology on the pre-operative and peri-operative samples. This research was funded by the Arthritis Research Campaign (grant number 16418).

Author information

Authors and Affiliations

  1. Infection and Immunity Research Group, Level 9, Glasgow Dental Hospital & School, 378 Sauchiehall Street, Glasgow, G2 3JZ, UK

    M. P. Riggio, Kate E. Dempsey, Allan Lennon, Gordon Ramage & Jeremy Bagg

  2. The Queen Elizabeth National Spinal Injuries Unit Scotland, South Glasgow University Hospitals Division, Southern General Hospital, 1345 Govan Road, Glasgow, G51 4TF, UK

    David Allan

Authors
  1. M. P. Riggio
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kate E. Dempsey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Allan Lennon
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. David Allan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gordon Ramage
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jeremy Bagg
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. P. Riggio.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Riggio, M.P., Dempsey, K.E., Lennon, A. et al. Molecular detection of transcriptionally active bacteria from failed prosthetic hip joints removed during revision arthroplasty. Eur J Clin Microbiol Infect Dis 29, 823–834 (2010). https://doi.org/10.1007/s10096-010-0934-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10096-010-0934-y

Keywords

Search

Navigation