The role of small-colony variants in failure to diagnose and treat biofilm infections in orthopedics

Neut, Daniëlle; Mei, Henny C. van der; Bulstra, Sjoerd K.; Busscher, Henk J.

doi:10.1080/17453670710013843

Cited by 114 publications

(102 citation statements)

References 74 publications

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“…Data regarding the effectiveness of scraping to dislodge bacteria from biofilm are scarce and contrasting. In one clinical study, scraping was better than conventional methods, such as culture of periprosthetic soft tissues, as a diagnostic tool [17]. In an in vitro study comparing scraping with sonication, the latter allowed the highest bacterial recovery [4].…”

Section: Discussionmentioning

confidence: 99%

Does Dithiothreitol Improve Bacterial Detection from Infected Prostheses? A Pilot Study

Drago

Romanò

Mattina

et al. 2012

Clinical Orthopaedics &Amp; Related Research

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Section: Discussionmentioning

confidence: 99%

Does Dithiothreitol Improve Bacterial Detection from Infected Prostheses? A Pilot Study

Drago

Romanò

Mattina

et al. 2012

Clinical Orthopaedics &Amp; Related Research

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“…This fact is in line with the notion that staphylococci and streptococci are the predominant causal agents in septic arthritis and bacterial endocarditis, but it also suggests that this test can be useful in the case of infections in which culture can be negative due to the ability of bacteria to grow in biofilms (Costerton et al, 2011;Stoodley et al, 2010). There are multiple possible reasons for the culture insensitivity of biofilm bacteria: a sufficient number of bacteria may not detach from the biofilm foci so as to reach the level of detectability by culture (Stoodley et al, 2010); repressed respiratory activities of biofilm bacteria (Nguyen et al, 2002), presence of S. aureus small colony variants that, in orthopaedic infections, has been identified as a possible explanation for the high rate of false negatives associated with conventional culture methods (Neut et al, 2007). Interestingly, Grampositive cocci were often evident on Gram staining of culture-negative specimens.…”

Section: Discussionmentioning

confidence: 99%

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

Mencacci

Leli

Cardaccia

et al. 2011

Journal of Medical Microbiology

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Early detection of aetiological agents is pivotal for adequate therapy for bacterial infections. Although culture is still considered the mainstay for laboratory diagnosis, it often lacks sensitivity, especially in patients already treated with antibiotics. The present study investigated the potential clinical utility of the commercial real-time-PCR-based system SeptiFast (SF), originally intended for diagnosis of sepsis from blood specimens, in the aetiological diagnosis of other bacterial infections, in patients undergoing antibiotic therapy. A total of 53 non-blood specimens were analysed for microbial pathogen detection by conventional culture and with SF real-time PCR: 19 (35.8 %) synovial fluids, 9 (17.0 %) cardiac valve tissues and 25 (47.2 %) purulent exudates from various body sites. Overall, the number of specimens positive for a pathogen by SF (26/53; 49.1 %) was significantly greater (P50.001) than that of specimens positive by culture (10/53; 18.9 %). In particular, SF was superior to culture for pathogen detection in cardiac valve tissues and synovial fluids. The analysis of concordance showed a fair agreement between the two methods (kappa value50.314; 95 % confidence interval50.531-0.097). Even with the limitation of the low number of specimens, this study confirmed the great potential of diagnosing bacterial infections by a molecular approach, and indicates that the real-time PCR SF system can be used for specimens other than blood, from patients undergoing antibiotic treatment.

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“…This tolerance may be conferred as a result of the decreased efficacy of antibiotics on bacterial persisters in the biofilm, developed antibiotic genetic resistance by the bacteria, or the extracellular polymeric substance preventing diffusion of the antibiotic through the biofilm [12,16].…”

Section: Introductionmentioning

confidence: 99%

Antibiotic-tolerant Staphylococcus aureus Biofilm Persists on Arthroplasty Materials

Urish

DeMuth

Kwan

et al. 2016

Clinical Orthopaedics &Amp; Related Research

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Background The continued presence of biofilm may be one cause of the high risk of failure observed with irrigation and débridement with component retention in acute periprosthetic joint infection (PJI). There is a poor understanding of the role of biofilm antibiotic tolerance in PJI. Questions/purposes (1) Do increasing doses of cefazolin result in decreased viable biofilm mass on arthroplasty materials? (2) Is cefazolin resistance phenotypic or genotypic? (3) Is biofilm viability a function of biofilm depth after treatment with cefazolin? (4) Is the toxin-antitoxin system, yoeB expression, associated with antibiotic stress?Methods Methicillin-sensitive Staphylococcus aureus biofilm was cultured on total knee arthroplasty (TKA) materials and exposed to increasing doses of cefazolin (control, 0.5, 1.0, 10.0, 100.0 lg/mL). Quantitative confocal microscopy and quantitative culture were used to measure viable biofilm cell density. To determine if cefazolin resistance was phenotypic or genotypic, we measured minimum inhibitory concentration (MIC) after exposure to different cefazolin concentrations; changes in MIC would suggest genotypic features, whereas unchanged MIC would suggest phenotypic behavior. Finally, quantitative reverse transcription-polymerase chain reaction was used to quantify expression of yoeB levels between biofilm and planktonic bacteria after exposure to 1 lg/mL cefazolin for 3 hours. Results Although live biofilm mass was reduced by exposure to cefazolin when compared with biofilm mass in controls (39.2 9 10 3 ± 26.4 9 10 3 pixels), where the level after 0.5 lg/mL exposure also showed reduced mass (20.3 9 10 3 ± 11.9 9 10 3 pixels), no further reduction was seen after higher doses (mass at 1.0 lg/mL: 5.0 9 10 3 pixels ± 1.1 9 10 3 pixels; at 10.0 lg/mL: 6.4 9 10 3 ± 9.6 9 10 3 pixels; at 100.0 lg/mL: 6.4 9 10 3 ± 3.9 9 10 3 ). At the highest concentration tested (100 lg/ mL), residual viable biofilm was present on all three materials, and there were no differences in percent biofilm survival among cobalt-chromium (18.5% ± 15.1%),The project described was supported by the National Institutes of Health through Grant Number KL2 TR000146 (KLU 123Clin Orthop Relat Res (2016) 474:1649-1656 DOI 10.1007 Clinical Orthopaedics and Related Research ® A Publication of The Association of Bone and Joint Surgeons® polymethylmethacrylate (22.8% ± 20.2%), and polyethylene (14.7% ± 10.4%). We found that tolerance was a phenotypic phenomenon, because increasing cefazolin exposure did not result in changes in MIC as compared with controls (MIC in controls: 0.13 ± 0.02; at 0.5 lg/mL: 0.13 ± 0.001, p = 0.96; at 1.0 lg/m: 0.14 ± 0.04, p = 0.95; at 10.0 lg/m: 0.11 ± 0.016, p = 0.47; at 100.0 lg/m: 0.94 ± 0.047, p = 0.47). Expression of yoeB after 1 lg/mL cefazolin for 3 hours in biofilm cells was greater in biofilm but not in planktonic cells (biofilm: 62.3-fold change, planktonic cells: À78.8-fold change, p \ 0.001). Conclusions Antibiotics are inadequate at complete removal of the biofilm from the surface o...

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The role of small-colony variants in failure to diagnose and treat biofilm infections in orthopedics

Cited by 114 publications

References 74 publications

Does Dithiothreitol Improve Bacterial Detection from Infected Prostheses? A Pilot Study

Does Dithiothreitol Improve Bacterial Detection from Infected Prostheses? A Pilot Study

Comparison of conventional culture with SeptiFast real-time PCR for microbial pathogen detection in clinical specimens other than blood

Antibiotic-tolerant Staphylococcus aureus Biofilm Persists on Arthroplasty Materials

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