PMID- 27749085 OWN - NLM STAT- MEDLINE DCOM- 20170404 LR - 20210910 IS - 1945-0257 (Electronic) IS - 1945-0265 (Print) IS - 1945-0257 (Linking) VI - 20 IP - 12 DP - 2016 Dec TI - Polymerase Chain Reaction-Electrospray-Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis. PG - 721-731 LID - 10.1089/gtmb.2016.0080 [doi] AB - BACKGROUND: Preliminary studies have identified known bacterial pathogens in the knees of patients with osteoarthritis (OA) before arthroplasty. AIMS: The current study was designed to determine the incidence and types of bacteria present in the synovial fluid of native knee joints from adult patients with diagnoses of septic arthritis and OA. PATIENTS AND METHODS: Patients were enrolled between October 2010 and January 2013. Synovial fluid samples from the affected knee were collected and evaluated with both traditional microbial culture and polymerase chain reaction-electrospray ionization-time-of-flight mass spectrometry (molecular diagnostics [MDx]) to prospectively characterize the microbial content. Patients were grouped by diagnosis into one of two cohorts, those with clinical suspicion of septic arthritis (n = 44) and those undergoing primary arthroplasty of the knee for OA (n = 21). In all cases where discrepant culture and MDx results were obtained, we performed species-specific 16S rRNA fluorescence in situ hybridization (FISH) as a confirmatory test. RESULTS: MDx testing identified bacteria in 50% of the suspected septic arthritis cases and 29% of the arthroplasty cases, whereas culture detected bacteria in only 16% of the former and 0% of the latter group. The overall difference in detection rates for culture and MDx was very highly significant, p-value = 2.384 x 10(-7). All of the culture-positive cases were typed as Staphylococcus aureus. Two of the septic arthritis cases were polymicrobial as was one of the OA cases by MDx. FISH testing of the specimens with discordant results supported the MDx findings in 91% (19/21) of the cases, including one case where culture detected S. aureus and MDx detected Streptococcus agalactiae. CONCLUSIONS: MDx were more sensitive than culture, as confirmed by FISH. FISH only identifies bacteria that are embedded or infiltrated within the tissue and is thus not susceptible to contamination. Not all suspected cases of septic arthritis contain bacteria, but a significant percent of patients with OA, and no signs of infection, have FISH-confirmed bacterial biofilms present in the knee. FAU - Palmer, Michael P AU - Palmer MP AD - 1 88th Surgical Operations Squadron, Orthopedic Surgery , Wright-Patterson Airforce Base, Dayton, Ohio. FAU - Melton-Kreft, Rachael AU - Melton-Kreft R AD - 2 Center of Excellence in Biofilm Research , Allegheny Health Network, Pittsburgh, Pennsylvania. FAU - Nistico, Laura AU - Nistico L AD - 2 Center of Excellence in Biofilm Research , Allegheny Health Network, Pittsburgh, Pennsylvania. FAU - Hiller, N Louisa AU - Hiller NL AD - 3 Department of Biological Sciences, Carnegie Mellon University , Pittsburgh, Pennsylvania. FAU - Kim, Leon H J AU - Kim LH AD - 3 Department of Biological Sciences, Carnegie Mellon University , Pittsburgh, Pennsylvania. FAU - Altman, Gregory T AU - Altman GT AD - 4 Department of Orthopaedic Surgery, Allegheny General Hospital , Pittsburgh, Pennsylvania. FAU - Altman, Daniel T AU - Altman DT AD - 4 Department of Orthopaedic Surgery, Allegheny General Hospital , Pittsburgh, Pennsylvania. FAU - Sotereanos, Nicholas G AU - Sotereanos NG AD - 4 Department of Orthopaedic Surgery, Allegheny General Hospital , Pittsburgh, Pennsylvania. FAU - Hu, Fen Z AU - Hu FZ AD - 5 Institute for Molecular Medicine and Infectious Disease, Center for Genomic Sciences, Drexel University College of Medicine , Philadelphia, Pennsylvania. FAU - De Meo, Patrick J AU - De Meo PJ AD - 4 Department of Orthopaedic Surgery, Allegheny General Hospital , Pittsburgh, Pennsylvania. FAU - Ehrlich, Garth D AU - Ehrlich GD AD - 5 Institute for Molecular Medicine and Infectious Disease, Center for Genomic Sciences, Drexel University College of Medicine , Philadelphia, Pennsylvania. AD - 6 Institute for Molecular Medicine and Infectious Disease, Center for Advanced Microbial Processing, Drexel University College of Medicine , Philadelphia, Pennsylvania. AD - 7 Department of Microbiology and Immunology, Drexel University College of Medicine , Philadelphia, Pennsylvania. AD - 8 Department of Otolaryngology-Head and Neck Surgery, Drexel University College of Medicine , Philadelphia, Pennsylvania. LA - eng GR - R01 AI080935/AI/NIAID NIH HHS/United States GR - R01 DC002148/DC/NIDCD NIH HHS/United States PT - Journal Article DEP - 20161017 PL - United States TA - Genet Test Mol Biomarkers JT - Genetic testing and molecular biomarkers JID - 101494210 RN - 0 (RNA, Ribosomal, 16S) SB - IM MH - Adult MH - Arthritis, Infectious/diagnosis/*microbiology MH - Bacterial Typing Techniques/methods MH - Female MH - Humans MH - In Situ Hybridization, Fluorescence MH - Male MH - Mass Spectrometry/methods MH - Molecular Diagnostic Techniques/methods MH - Osteoarthritis, Knee/diagnosis/*microbiology MH - Polymerase Chain Reaction/methods MH - RNA, Ribosomal, 16S/genetics MH - Staphylococcus aureus/classification/genetics/*isolation & purification MH - Synovial Fluid/microbiology PMC - PMC5180073 OTO - NOTNLM OT - bacterial infection OT - fluorescent in situ hybridization OT - molecular diagnostics OT - osteoarthritis OT - septic arthritis COIS- Author Disclosure Statement No competing financial interests exist. EDAT- 2016/10/18 06:00 MHDA- 2017/04/05 06:00 PMCR- 2016/12/01 CRDT- 2016/10/18 06:00 PHST- 2016/10/18 06:00 [pubmed] PHST- 2017/04/05 06:00 [medline] PHST- 2016/10/18 06:00 [entrez] PHST- 2016/12/01 00:00 [pmc-release] AID - 10.1089/gtmb.2016.0080 [pii] AID - 10.1089/gtmb.2016.0080 [doi] PST - ppublish SO - Genet Test Mol Biomarkers. 2016 Dec;20(12):721-731. doi: 10.1089/gtmb.2016.0080. Epub 2016 Oct 17.