Role of Dithiothreitol in Detection of Orthopaedic Implant-Associated Infections.

Orthopaedic implant-associated infections (OIAIs) represent a notable complication of contemporary surgical procedures, exerting a considerable impact on patient outcomes and escalating healthcare expenditures. Prompt diagnosis holds paramount importance in managing OIAIs, with sonication widely acknowledged as the preferred method for detecting biofilm-associated infections. Recently, dithiothreitol (DTT) has emerged as a potential substitute for sonication, owing to its demonstrated ability to impede biofilm formation. This study aimed to compare the efficacy of DTT with sonication in identifying microorganisms within implants. Conducted as a prospective cohort investigation, the study encompassed two distinct groups: patients with suspected infections undergoing implant removal (Group A) and those slated for hardware explantation (Group B). Hardware segments were assessed for biofilm-related microorganisms using both sonication and DTT, with a comparative analysis of the two methods. A total of 115 patients were enrolled. In Group A, no statistically significant disparity was observed between DTT and sonication. DTT exhibited a sensitivity of 89.47% and specificity of 96.3%. Conversely, in Group B, both DTT and sonication fluid cultures yielded negative results in all patients. Consequently, this investigation suggests that DTT holds comparable efficacy to sonication in detecting OIAIs, offering a novel, cost-effective, and readily accessible diagnostic modality for identifying implant-associated infections.

Sonication of revised hip and knee prostheses detects occult infections, improves clinical outcomes and prevents re - revisions. A case series study.

Introduction: Periprosthetic joint infection (PJI) is a devastating complication occurring in 1-2% of primary and up to 10% of revised total hip and knee arthroplasties (THA and TKA) impairing patient's quality of life. Occult infections are underdiagnosed, sub-treated and sub-clinically experienced by patients. This study aimed to correlate patients' clinical outcomes with early antibiotic treatment based on use or non-use of a sonication technique on explanted prostheses. Methods: 33 patients with revised THA or TKA were retrospectively evaluated. Clinical outcomes were assessed via Oxford hip or knee scores, and correlated with administration or not of antibiotic treatment based on sonication results. Results: According to laboratory findings the patients were divided in the following three groups: 1. Septic loosening (conventional cultures and/or sonication positive), 2. Aseptic loosening (conventional cultures and sonication negative) and 3. Occult loosening (conventional cultures negative, sonication not performed). The average Oxford score was poor (27.9/60) for the septic, excellent (43.8/60) for the aseptic and intermediate (37.7/60) for the occult group. Additionally, conventional cultures were negative, but sonication-positive, in 6 individuals with patient-related risk factors (male gender, BMI > 30 kg/m2, diabetes, hypertension, steroids and rheumatoid arthritis). Conclusions: Sonication represents a valuable diagnostic technique to guide administration of effective antibiotic treatment for patients, especially for detection of persistent post-revision occult infections. We recommend the systematic investigation of revised prostheses with a sonication technique, but especially in cases with risk factors for infection who it is suspected may have occult loosening.

A comparative study of heterogeneous antibiotic resistance of microbial populations in conventional periprosthetic tissue cultures and sonication fluid cultures of orthopaedics explanted prostheses.

BACKGROUND: "Heterogeneity" describes a phenomenon where subpopulations of seemingly isogenic bacteria exhibit a range of susceptibilities to a particular antibiotic. We aim to investigate the frequency of heterogeneity among microbes isolated from infected prostheses, and its possible correlation with microbial resistance. METHODS: Between May 2014 and June 2019, we investigated 234 patients, at our institution, undergoing revision arthroplasty because of loosening of the prostheses or because of periprosthetic joint infection. All patients had periprosthetic tissue culture, sonication of prosthesis and direct inoculation of Sonication fluid into blood culture bottles. We assessed the presence of heterogeneity among all pathogens isolated from infected prostheses. RESULTS: Using standard non-microbiological criteria to determine periprosthetic joint infection, it was found that 143 patient (61.1%) had aseptic loosening while 91 patients (38.9%) had periprosthetic joint infection. Comparing the two methods, the results of our study showed that the method of sonication was significantly more sensitive than tissue culture [91% (83-96) vs. 43% (33-54); p < 0.005]. In this study, heterogeneity was reported in 15 cases, 16.5% of all infections and 6.4% in the total population. In our study, Staphylococcus epidermidis was the most commonly isolated strain followed by Staphylococcus aureus, at a rate of 35.2% and 19.8%, respectively. Antibiotics in which the microorganisms exhibited heterogeneous bacterial behavior most frequently were Gendamicin (5.3%), Vancomycin (4.9%). CONCLUSION: There is increasing evidence that heterogeneity can lead to therapeutic failure and that the detection of this phenotype is a prerequisite for a proper antibiotic choice to have a successful therapeutic effect.

Combination of conventional culture, vial culture, and broad-range PCR of sonication fluid for the diagnosis of prosthetic joint infection.

We assessed the value of conventional culture, vial culture, and broad-range PCR of the sonication fluid (SF), individually or in combinations, for the diagnosis of prosthetic joint infection (PJI). We studied 114 consecutive patients (median age:72.5 years, males: 28.07%) undergoing removal of a total knee or hip prosthesis. By non-microbiologic criteria, 87 patients had aseptic failure, and 27 PJI. All patients had periprosthetic tissue culture, sonication of prosthesis, and study of SF by conventional and vial culture, and PCR. Compared to tissue culture, each test was significantly more sensitive and less specific. If only one test was positive, the sensitivity was 88.46% and specificity 64.29%. If all three SF tests were positive, sensitivity, and NPV were decreasing (34.6% and 80.23%), but specificity and PPV were increasing up to 98.57% and 90.9%, respectively, outperforming tissue culture. A triple negative test practically excluded PJI.

Superiority of the sonication method against conventional periprosthetic tissue cultures for diagnosis of prosthetic joint infections.

INTRODUCTION: Diagnosis of periprosthetic infections is challenging. The aim of this study was to compare the diagnostic accuracy of conventional periprosthetic tissue culture and culture of sonication fluid of the explanted prostheses. METHODS: We prospectively enrolled 114 patients undergoing revision hip or knee arthroplasty because of loosening of the prostheses, at our institution, between July 2012 and July 2016. Patients' medical history and demographic characteristics were recorded. The explanted hardware was separated in sterile containers and sonicated under sterile conditions. At least five samples of periprosthetic tissue were sent for culture and histological examination. We compared the culture of samples obtained by sonication of explanted hip and knee prostheses with conventional culture of periprosthetic tissue for the microbiological diagnosis of prosthetic joint infection. RESULTS: Infectious Diseases Society of America guidelines were used for the definition of prosthetic joint infection. Sixty-one patients had periprosthetic infection and 53 aseptic loosening (73 hip prostheses and 41 knee prostheses). The sensitivity of sonication fluid culture was 77.04%, and the sensitivity of conventional tissue cultures was 55.73% (p value = 0.012). The specificities of the two methods were 98.11 and 94.34%, respectively. The sensitivity of the histopathological examination of the periprosthetic tissue was 72.10%. There were 17 patients with PJI where the isolated pathogen was detected in SFC but not in PTC, while in five cases the pathogen was detected only in PTC. There were nine patients where no bacteria were detected by any microbiological method and the diagnosis was based on clinical and histological findings, according to the guidelines. CONCLUSIONS: The sonication method represents a reliable test for the diagnosis of prosthetic joint infections with a greater sensitivity and specificity than the conventional periprosthetic tissue cultures.

Point-prevalence survey of healthcare facility-onset healthcare-associated Clostridium difficile infection in Greek hospitals outside the intensive care unit: The C. DEFINE study.

BACKGROUND: The correlation of Clostridium difficile infection (CDI) with in-hospital morbidity is important in hospital settings where broad-spectrum antimicrobial agents are routinely used, such as in Greece. The C. DEFINE study aimed to assess point-prevalence of CDI in Greece during two study periods in 2013. METHODS: There were two study periods consisting of a single day in March and another in October 2013. Stool samples from all patients hospitalized outside the ICU aged ≥18 years old with diarrhea on each day in 21 and 25 hospitals, respectively, were tested for CDI. Samples were tested for the presence of glutamate dehydrogenase antigen (GDH) and toxins A/B of C. difficile; samples positive for GDH and negative for toxins were further tested by culture and PCR for the presence of toxin genes. An analysis was performed to identify potential risk factors for CDI among patients with diarrhea. RESULTS: 5,536 and 6,523 patients were screened during the first and second study periods, respectively. The respective point-prevalence of CDI in all patients was 5.6 and 3.9 per 10,000 patient bed-days whereas the proportion of CDI among patients with diarrhea was 17% and 14.3%. Logistic regression analysis revealed that solid tumor malignancy [odds ratio (OR) 2.69, 95% confidence interval (CI): 1.18-6.15, p = 0.019] and antimicrobial administration (OR 3.61, 95% CI: 1.03-12.76, p = 0.045) were independent risk factors for CDI development. Charlson's Comorbidity Index (CCI) >6 was also found as a risk factor of marginal statistical significance (OR 2.24, 95% CI: 0.98-5.10). Median time to CDI from hospital admission was shorter with the presence of solid tumor malignancy (3 vs 5 days; p = 0.002) and of CCI >6 (4 vs 6 days, p = 0.009). CONCLUSIONS: The point-prevalence of CDI in Greek hospitals was consistent among cases of diarrhea over a 6-month period. Major risk factors were antimicrobial use, solid tumor malignancy and a CCI score >6.

Sonication assisted microbiological diagnosis of implant-related infection caused by Prevotella disiens and Staphylococcus epidermidis in a patient with cranioplasty.

BACKGROUND: Infections present a major complication of cranioplasty procedures and in many cases removal of the implant material becomes a necessity. Sonication of the artificial implant material has been used during the last years, in order to facilitate better diagnosis of these infections, nevertheless its use in cranial implant infections is still limited. CASE PRESENTATION: A case of a 63-year-old Caucasian male patient who underwent a decompressive craniectomy, due to intracranial hemorrhage, and a consequent cranioplasty using an autogenic bone flap fixed by titanium clamps, is reported. After three unsuccessful cranioplasty efforts to repair a persistent skin defect, removing the bone flap and the titanium clamps was a necessity. Tissue and bone cultures were unable to reveal any microorganism whilst sonication of the removed titanium clamps and consequent culture of the resulting sonication liquid yielded Prevotella disiens and Staphylococcus epidermidis. The patient was treated with daptomycin and metronidazole until discharge and the skin defect was successfully repaired. CONCLUSION: The present case report indicates that the use of the sonication procedure assisted the microbiological diagnosis. This is the first known neurosurgical case of the implementation of the sonication procedure.

Wide dissemination of linezolid-resistant Staphylococcus epidermidis in Greece is associated with a linezolid-dependent ST22 clone.

OBJECTIVES: Dependence on linezolid was recently described as significant growth acceleration of linezolid-resistant Staphylococcus epidermidis (LRSE) isolates upon linezolid exposure. We investigated the possible contribution of linezolid dependence to LRSE dissemination in Greece. METHODS: Linezolid resistance rates were estimated in six tertiary hospitals located throughout Greece between 2011 and 2013. Sixty-three randomly selected LRSE recovered in these hospitals during this period were studied. Growth curve analysis was conducted with and without linezolid. Clonality of the isolates was investigated by PFGE and MLST. RESULTS: During the study period, the LRSE rate in the participating hospitals rose significantly from 6.9% to 9% (P = 0.006); the increase was more prominent in ICUs (from 15.1% to 20.9%; P = 0.005). Forty-seven (74.6%) of the 63 LRSE, derived from all study hospitals, clearly exhibited linezolid dependence, growing significantly faster in the presence of 16 and 32 mg/L linezolid. Of note, 61 (96.8%) LRSE exhibited a single macrorestriction pattern and belonged to ST22, which included all linezolid-dependent LRSE. The remaining two LRSE belonged to unique STs. Five of six linezolid-dependent isolates tested also exhibited linezolid dependence upon exposure to 8 mg/L linezolid. Interestingly, five of six ST22 linezolid-non-dependent isolates tested developed linezolid dependence when linezolid exposure preceded growth analysis. CONCLUSIONS: The rapid LRSE dissemination in Greek hospitals threatens linezolid activity. The observation that most LRSE belonged to ST22 and expressed dependence on linezolid clearly implies that the spread of linezolid resistance should have been driven by this trait, which provided the LRSE with a selective advantage under linezolid pressure.