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Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis

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 patien...

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Autores principales: Palmer, Michael P., Melton-Kreft, Rachael, Nistico, Laura, Hiller, N. Louisa, Kim, Leon H.J., Altman, Gregory T., Altman, Daniel T., Sotereanos, Nicholas G., Hu, Fen Z., De Meo, Patrick J., Ehrlich, Garth D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5180073/
https://www.ncbi.nlm.nih.gov/pubmed/27749085
http://dx.doi.org/10.1089/gtmb.2016.0080
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author Palmer, Michael P.
Melton-Kreft, Rachael
Nistico, Laura
Hiller, N. Louisa
Kim, Leon H.J.
Altman, Gregory T.
Altman, Daniel T.
Sotereanos, Nicholas G.
Hu, Fen Z.
De Meo, Patrick J.
Ehrlich, Garth D.
author_facet Palmer, Michael P.
Melton-Kreft, Rachael
Nistico, Laura
Hiller, N. Louisa
Kim, Leon H.J.
Altman, Gregory T.
Altman, Daniel T.
Sotereanos, Nicholas G.
Hu, Fen Z.
De Meo, Patrick J.
Ehrlich, Garth D.
author_sort Palmer, Michael P.
collection PubMed
description 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 × 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.
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spelling pubmed-51800732017-01-11 Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis Palmer, Michael P. Melton-Kreft, Rachael Nistico, Laura Hiller, N. Louisa Kim, Leon H.J. Altman, Gregory T. Altman, Daniel T. Sotereanos, Nicholas G. Hu, Fen Z. De Meo, Patrick J. Ehrlich, Garth D. Genet Test Mol Biomarkers Original Articles 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 × 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. Mary Ann Liebert, Inc. 2016-12-01 2016-12-01 /pmc/articles/PMC5180073/ /pubmed/27749085 http://dx.doi.org/10.1089/gtmb.2016.0080 Text en © Michael P. Palmer et al., 2016; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons Attribution Noncommercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited.
spellingShingle Original Articles
Palmer, Michael P.
Melton-Kreft, Rachael
Nistico, Laura
Hiller, N. Louisa
Kim, Leon H.J.
Altman, Gregory T.
Altman, Daniel T.
Sotereanos, Nicholas G.
Hu, Fen Z.
De Meo, Patrick J.
Ehrlich, Garth D.
Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title_full Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title_fullStr Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title_full_unstemmed Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title_short Polymerase Chain Reaction–Electrospray–Time-of-Flight Mass Spectrometry Versus Culture for Bacterial Detection in Septic Arthritis and Osteoarthritis
title_sort polymerase chain reaction–electrospray–time-of-flight mass spectrometry versus culture for bacterial detection in septic arthritis and osteoarthritis
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5180073/
https://www.ncbi.nlm.nih.gov/pubmed/27749085
http://dx.doi.org/10.1089/gtmb.2016.0080
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