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Journal of Clinical Microbiology, October 1999, p. 3281-3290, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Prosthetic Hip Infection at Revision Arthroplasty by Immunofluorescence Microscopy and PCR Amplification of the Bacterial 16S rRNA Gene

Michael M. Tunney,1 Sheila Patrick,1,* Martin D. Curran,3 Gordon Ramage,1 Donna Hanna,1 James R. Nixon,4 Sean P. Gorman,2 Richard I. Davis,5 and Neil Anderson5

Department of Microbiology and Immunobiology, School of Clinical Medicine, The Queen's University of Belfast, Belfast BT12 6BN,1 Regional Histocompatibility and Immunogenetics Laboratory, Belfast City Hospital, Belfast BT9 7TS,3 Withers Orthopaedic Centre, Musgrave Park Hospital, Belfast BT9 7JB,4 School of Pharmacy, The Queen's University of Belfast, Belfast BT9 7BL,2 and Department of Pathology, The Royal Group of Hospitals and Dental Hospital Health and Social Services Trust, Belfast BT12 6BA,5 United Kingdom

Received 19 January 1999/Returned for modification 2 May 1999/Accepted 26 June 1999

In this study the detection rates of bacterial infection of hip prostheses by culture and nonculture methods were compared for 120 patients with total hip revision surgery. By use of strict anaerobic bacteriological practice during the processing of samples and without enrichment, the incidence of infection by culture of material dislodged from retrieved prostheses after ultrasonication (sonicate) was 22%. Bacteria were observed by immunofluorescence microscopy in 63% of sonicate samples with a monoclonal antibody specific for Propionibacterium acnes and polyclonal antiserum specific for Staphylococcus spp. The bacteria were present either as single cells or in aggregates of up to 300 bacterial cells. These aggregates were not observed without sonication to dislodge the biofilm. Bacteria were observed in all of the culture-positive samples, and in some cases in which only one type of bacterium was identified by culture, both coccoid and coryneform bacteria were observed by immunofluorescence microscopy. Bacteria from skin-flake contamination were readily distinguishable from infecting bacteria by immunofluorescence microscopy. Examination of skin scrapings did not reveal large aggregates of bacteria but did reveal skin cells. These were not observed in the sonicates. Bacterial DNA was detected in 72% of sonicate samples by PCR amplification of a region of the bacterial 16S rRNA gene with universal primers. All of the culture-positive samples were also positive for bacterial DNA. Evidence of high-level infiltration either of neutrophils or of lymphocytes or macrophages into associated tissue was observed in 73% of patients. Our results indicate that the incidence of prosthetic joint infection is grossly underestimated by current culture detection methods. It is therefore imperative that current clinical practice with regard to the detection and subsequent treatment of prosthetic joint infection be reassessed in the light of these results.

* Corresponding author. Mailing address: Department of Microbiology and Immunobiology, School of Clinical Medicine, The Queen's University of Belfast, Grosvenor Rd., Belfast BT12 6BN, United Kingdom. Phone: 01232-263205. Fax: 01232-439181. E-mail: s.patrick{at}qub.ac.uk.

Journal of Clinical Microbiology, October 1999, p. 3281-3290, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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