THE USE OF A REAL TIME PCR FOR DIAGNOSIS OF HIP AND KNEE PROSTHETIC INFECTION: METHOD DEVELOPMENT AND COMPARISON WITH HISTOLOGY AND BACTERIAL CULTURE

Abstract

Introduction: etiology of late infection after arthroplasty can be difficult to establish. Histology is the gold standard for infection in patients without inflammatory arthritis but diagnosis in inflammatory arthritis depends on culture (Atkins et al). Real-time PCR offers a rapid and direct assessment for staphylococci and enterococci infection but has not been widely assessed.

The aims of this study were

1. to develop the Roche lightcycler Staphylococcal and Enterococcal PCR kits to facilitate diagnosis of hip and knee prosthetic infections

2. To analyse results together with bacteriological and histological findings.

Methods: uplicate, multiple tissue samples were taken (with separate sterile instruments) at the 1^st stage of revision after informed consent. One set were cultured and results interpreted by the Oxford criteria. The second set were extracted using the Qiagen DNA kit, purified (in-house method) and tested using the Roche lightcycler kits.

Results:53 patients undergoing 2 stage revision for suspected infection were recruited.15 (28.3%) had negative histology and no inflammatory arthritis; 3 with single positive cultures and negative PCR – considered contaminants.

29 patients had non-inflammatory arthritis. 14/18 (77.8%) with positive cultures had staphylococci +/or enterococci isolated and 10 PCR results correlated. The other 11 patients had negative cultures.

9 patients had inflammatory arthritis. Six were culture negative and of the other three, 2 were positive for staphylococci on culture with 1 positive by PCR.

Discussion: Negative staphylococcal PCR correlates with the isolation of staphylococci from only one sample. This agrees with the Oxford criteria that such samples may be considered contaminants. Additional positives detected by staphylococcal PCR alone are rare.

Enterococcal PCR confirmed culture positivity in 2/3 patients. An additional 5 positive PCR’s were obtained from patients’ culture negative for enterococci. It is not clear if these are false positives or more sensitive detection of enterococcal isolation.