Skip to main content
SpringerLink
Log in

Sponge swabs increase sensitivity of sterility testing of processed bone and tendon allografts

  • Short Communication
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Sterility testing is the final, and critical, step in quality control of tissue banking. It informs the decision whether to release the tissue allografts for clinical use, or not. The most common method for sterility testing of structural bone and tendon allografts is to swab using cotton tip streaks. This method provides low recovery efficiency; and therefore may pass allografts with low bioburden, providing false negatives. Our pilot data revealed organism recovery efficiencies of 60, 30 and 100% from cotton swab, membrane filtration and sponge swaps, respectively. Our aim was to develop a high sensitivity sterility test for structural bone and tendon allografts using a sponge sampling method. Eighty-one bone and tendon allograft samples were inoculated with organism suspensions (102 or less organisms per 0.1 mL) of Clostridium sporogenes, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, Bacillus subtilis, Aspergillus niger, Staphylococcus epidermidis and Micrococcus spp. Nasco sponges (4 × 8 cm) were used to aseptically sample the whole surface of allograft samples. The sponges were cut in half and cultured in either tryptone soya or fluid thioglycollate broths for 14 days. Positive culture samples were further examined for microbial morphology. The results showed that the sensitivity of the method, and negative predictive value, is 100% for all inoculated organisms incubated with thioglycollate. We conclude that this sponge sampling method should be applied as the standard for sterility testing of structural bone and tendon allografts.

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

References

  1. American Association of Tissue Banks (AATB) (2002) Standards for tissue banking, 10th edn. AATB, McLean

    Google Scholar 

  2. Barrios RH, Leyes M, Amillo S, Oteiza C (1994) Bacterial contamination of allografts. Acta Orthop Belg 60(2):152–154

    PubMed  CAS  Google Scholar 

  3. Byrne B, Dunne G, Lyng J, Bolton DJ (2005) Microbiological carcass sampling methods to achieve compliance with 2001/471/EC and new hygiene regulations. Res Microbiol 156(1):104–106. doi:10.1016/j.resmic.2004.08.007

    Article  PubMed  Google Scholar 

  4. Chiu CK, Lau PY, Chan SWW, Fong CM, Sun LK (2004) Microbial contamination of femoral head allografts. Hong Kong Med J 10(6):401–405

    PubMed  CAS  Google Scholar 

  5. Dennis J, Martinez O, Landy D, Malinin T, Morris P, Fox W, Buck B, Temple H (2011) A comparison of two microbial detection methods used in aseptic processing of musculoskeletal allograft tissues. Cell Tissue Bank 12:45–50

    Google Scholar 

  6. Eblen DR, Levine P, Rose BE, Saini P, Mageau R, Hill WE (2005) Nationwide microbiological baseline data collected by sponge sampling during 1997 and 1998 for cattle, swine, turkeys, and geese. J Food Prot 68(9):1848–1852

    PubMed  CAS  Google Scholar 

  7. Gill CO, Badoni M (2010) Effects of experience with swabbing procedures on the numbers of bacteria recovered from carcasses by swabbing with sponges. J Food Prot 73(4):747–751

    PubMed  CAS  Google Scholar 

  8. Gill CO, Badoni M, McGinnis JC (2001) Microbiological sampling of meat cuts and manufacturing beef by excision or swabbing. J Food Prot 64(3):325–334

    PubMed  CAS  Google Scholar 

  9. Husted H, Kramhoft MU (1996) Microbiology of femoral head grafts in bone banks. Ugeskr Laeger 158(44):6260–6262

    PubMed  CAS  Google Scholar 

  10. Ibrahim T, Stafford H, Esler CNA, Power RA (2004) Cadaveric allograft microbiology. Int Orthop 28(5):315–318. doi:10.1007/s00264-004-0579-5

    Article  PubMed  CAS  Google Scholar 

  11. International Standard Organisation (2006) Annex B–guidance on methods of determination of bioburden. In: Technical Committee ISO/TC 198 (ed) ISO 11737–1: sterilisation of medical devices–microbiological methods. Part 1: determination of a population of microorganisms on products. ISO, Geneva, pp 22–30

    Google Scholar 

  12. International Standard Organisation (ISO) (2006) Annex A: guidance on determination of a production of microorganisms on product. In: ISO (ed) ISO 11737–1: sterilisation of medical devices–microbiological methods. Part 1: determination of a population of microorganisms on products. ISO, Geneva, pp 9–21

    Google Scholar 

  13. Ireland L, Spelman D (2005) Bacterial contamination of tissue allografts–experiences of the Donor Tissue Bank of Victoria. Cell Tissue Bank 6(3):181–189

    Article  PubMed  Google Scholar 

  14. Lee EH, Ferguson D, Jernigan D, Greenwald M, Cote T, Bos JE, Guarner J, Zaki S, Schuchat A, Beall B, Srinivasan A (2007) Invasive group-A streptococcal infection in an allograft recipient–A case report. J Bone Joint Surg Am 89A(9):2044–2047. doi:10.2106/jbjs.f.01594

    Article  Google Scholar 

  15. Lewandowski R, Kozlowska K, Szpakowska M, Stepinska M, Trafny EA (2010) Use of a foam spatula for sampling surfaces after bioaerosol deposition. Appl Environ Microbiol 76(3):688–694. doi:10.1128/aem.01849-09

    Article  PubMed  CAS  Google Scholar 

  16. Martinez B, Celda MF, Anastasio B, Garcia I, Lopez-Mendoza M (2010) Microbiological sampling of carcasses by excision or swabbing with three types of sponge or gauze. J Food Prot 73(1):81–87

    PubMed  Google Scholar 

  17. Maso A, Andollina A, Bassi A, Bertoni G, Modelli L, Quinto C, Vaccari M, Fornasari PM (2003) Bacterial contamination of musculoskeletal allografts. Chir Organi Mov 88(4):345–350

    PubMed  CAS  Google Scholar 

  18. Nguyen H, Morgan DA, Forwood MR (2011) Validation of 11 kGy as a radiation sterilization dose for frozen bone allografts. J Arthroplast 26(2):303–308. doi:10.1016/j.arth.2010.03.032

    Article  Google Scholar 

  19. Nguyen H, Morgan DAF, Sly LI, Benkovich M, Cull S, Forwood MR (2008) Validation of 15 kGy as a radiation sterilisation dose for bone allografts manufactured at the Queensland Bone Bank: application of the VDmax 15 method. Cell Tissue Bank 9(2):139–147

    Article  PubMed  CAS  Google Scholar 

  20. Ronholdt CJ, Bogdansky S (2005) The appropriateness of swab cultures for the release of human allograft tissue. J Ind Microbiol Biotechnol 32(8):349–354. doi:10.1007/s10295-005-0251-0

    Article  PubMed  CAS  Google Scholar 

  21. Saegeman VSM, Lismont D, Verduyckt B, Ectors NL, Verhaegen J (2007) Comparison of microbiological culture methods in screening allograft tissue. Swab versus nutrient broth. J Microbiol Methods 70(2):374–378

    Article  PubMed  CAS  Google Scholar 

  22. Sommerville SMM, Johnson N, Bryce SL, Journeaux SF, Morgan DAF (2000) Contamination of banked femoral head allograft: incidence, bacteriology and donor follow up. Aust N Z J Surg 70(7):480–484

    Article  PubMed  CAS  Google Scholar 

  23. Therapeutic Goods Administration (TGA) (2006) TGA guidelines for sterility testing of therapeutic goods. TGA, Canberra, p 9

  24. Therapeutic Goods Administration (TGA) (2006) TGA guidelines for sterility testing of therapeutic goods. TGA, Canberra

  25. United States Pharmacopeia (2009) Validation of microbial recovery from pharmacopeial articles. In: The United States Pharmacopeia, 32nd rev and the National Formulary, 27th edn (USP 32-NF 27). United States Pharmacopeia, Rockville, p 737

  26. Varettas K, Taylor P (2011) Bioburden assessment of banked bone used for allografts. Cell Tissue Bank (2011) 12:37–43

    Google Scholar 

  27. Veen MR, Bloem RM, Petit PLC (1994) Sensitivity and negative predictive values of swab culture in musculoskeletal allograft procurement. Clin Orthop Relat Res 300:259–263

    Google Scholar 

  28. Vehmeyer SBW, Bloem RM, Petit PLC (2001) Microbiological screening of post-mortem bone donors–two case reports. J Hosp Infect 47(3):193–197. doi:10.1053/jhin.2000.0836

    Article  PubMed  CAS  Google Scholar 

  29. Vehmeyer SBW, Slooff ARM, Bloem RM, Petit PLC (2002) Bacterial contamination of femoral head allografts from living donors. Acta Orthop Scand 73(2):165–169

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Queensland Bone Bank, NH & MRC Project Grant 453624, Arthritis Australia Project Grant and Griffith University Encouragement Grant. We thank Naomi Ross and Biotest Laboratories Pty Ltd, Queensland, Australia for technical expertise in microbiological assays.

Author information

Authors and Affiliations

  1. The Queensland Bone Bank, Brisbane, Queensland, Australia

    Huynh Nguyen, David A. F. Morgan, Sharon Cull & Morris Benkovich

  2. Brisbane Private Hospital, Brisbane, Queensland, Australia

    David A. F. Morgan

  3. School of Medical Science, Griffith University, Brisbane, Queensland, 4222, Australia

    Huynh Nguyen & Mark R. Forwood

  4. Musculoskeletal Research Program, Griffith Health Institute, Griffith University, Brisbane, Queensland, 4222, Australia

    Huynh Nguyen & Mark R. Forwood

Authors
  1. Huynh Nguyen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David A. F. Morgan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sharon Cull
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Morris Benkovich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mark R. Forwood
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mark R. Forwood.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nguyen, H., Morgan, D.A.F., Cull, S. et al. Sponge swabs increase sensitivity of sterility testing of processed bone and tendon allografts. J Ind Microbiol Biotechnol 38, 1127–1132 (2011). https://doi.org/10.1007/s10295-011-0982-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10295-011-0982-z

Keywords

Search

Navigation