Molecular diagnosis of periprosthetic joint infection by quantitative RT-PCR of bacterial 16S ribosomal RNA.
The diagnosis of periprosthetic joint infection is sometimes straightforward with purulent discharge from the fistula tract communicating to the joint prosthesis. However it is often difficult to differentiate septic from aseptic loosening of prosthesis because of the high culture-negative rates in conventional microbiologic culture. This study used quantitative reverse transcription polymerase chain reaction (RT-qPCR) to amplify bacterial 16S ribosomal RNA in vitro and in 11 clinical samples. The in vitro analysis demonstrated that the RT-qPCR method was highly sensitive with the detection limit of bacterial 16S rRNA being 0.148 pg/ μ l. Clinical specimens were analyzed using the same protocol. The RT-qPCR was positive for bacterial detection in 8 culture-positive cases (including aerobic, anaerobic, and mycobacteria) and 2 culture-negative cases. It was negative in one case that the final diagnosis was confirmed without infection. The molecular diagnosis of bacterial infection using RT-qPCR to detect bacterial 16S rRNA around a prosthesis correlated well with the clinical findings. Based on the promising clinical results, we were attempting to differentiate bacterial species or drug-resistant strains by using species-specific primers and to detect the persistence of bacteria during the interim period before the second stage reimplantation in a larger scale of clinical subjects.
Department of Orthopaedic Surgery, Chia-Yi Chang Gung Memorial Hospital, 6 West Sec., Chiapu Road, Putzu City, Chiayi County 613, Taiwan ; Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan ; College of Medicine, Chang Gung University, Putzu City, Chiayi County 613, Taiwan.
Mel S
MS
Lee
Department of Power Mechanical Engineering, National Tsing Hua University, No. 101, Section 2, Kuang-Fu Road, Hsinchu 30013, Taiwan.
Wen-Hsin
WH
Chang
Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan.
Su-Chin
SC
Chen
Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan.
Pang-Hsin
PH
Hsieh
Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan.
Hsin-Nung
HN
Shih
Department of Orthopaedic Surgery, Linkou Chang Gung Memorial Hospital, Putzu City, Chiayi County 613, Taiwan.
Steve W N
SW
Ueng
Department of Power Mechanical Engineering, National Tsing Hua University, No. 101, Section 2, Kuang-Fu Road, Hsinchu 30013, Taiwan ; Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Taiwan.
Gwo-Bin
GB
Lee
D000328:Adult; D000368:Aged; D001419:Bacteria / Q000235:genetics*; D001424:Bacterial Infections / Q000175:diagnosis / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004866:Equipment Contamination; D005260:Female; D006801:Humans; D007595:Joint Prosthesis; D008297:Male; D008875:Middle Aged; D012329:RNA, Bacterial / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S