Development of a molecular methodology to quantify Staphylococcus epidermidis in surgical wash-out samples from prosthetic joint replacement surgery
- نوع فایل : کتاب
- زبان : انگلیسی
- مؤلف : Fergus J. Byrne · Sinéad M. Waters · Peadar S. Waters ·William Curtin · Michael Kerin
- چاپ و سال / کشور: 2007
Description
Prosthetic joint infection is a serious complication of total joint arthroplasty that causes great morbidity in aVected individuals. The most common cause of prosthesis associated infections are members of Staphylococcus spp., including Staphylococcus epidermidis. Culture has served as the gold standard for diagnosis, despite obvious shortcomings in terms of sensitivity and time. Bacterial genomic DNA extraction methodologies were evaluated for optimal recovery of genomic DNA from sterilised wash-out samples, spiked with S. epidermidis. Real time Polymerase chain reaction (PCR) assays targeting the S. epidermidis speciWc gseA gene were designed to reliably detect and quantify S. epidermidis. Sixty post-operative wash-out samples from primary hip and knee arthroplasties were taken aseptically. All were shown to be culture negative using the culture-dependent approach. These were samples were subjected to S. epidermidis-speciWc real time PCR. Standard curve showed good linearity. Sensitivity limit of the assay was <10 CFU S. epidermidis per sample. Reproducibility of the assay was conWrmed. S. epidermidis was not identiWed in any of these samples using the novel species speciWc SYBR Green real time PCR technique. Results indicated that wash-out samples were true negatives and did not harbour S. epidermidis. To support this, patients displayed no symptoms of infection. To illustrate the full eVectiveness of the novel real time PCR assay, a larger number of samples need to be tested (>1,000 patients).
Eur J Orthop Surg Traumatol DOI 10.1007/s00590-007-0221-5 Received: 4 December 2006 / Accepted: 11 January 2007
