Antibiotic Resistance Patterns Among Uropathogens in Female Outpatients Affected by Uncomplicated Cystitis: Focus on Fosfomycin Trometamol.

OBJECTIVES: To report the resistance rate against fosfomycin trometamol among outpatient women with symptoms related to urinary tract infections over a 6-year period in a multicentre, cross-sectional study. METHODS: Urinary samples were collected from three high-volume laboratories from January 2015 to December 2020. The pattern of resistance to fosfomycin was analysed by using the Vitek II automated system. RESULTS: A total of 7289 urinary samples were collected and 8321 strains were analysed during the study period. The most commonly isolated uropathogen was Escherichia coli (n = 6583, 79.1%). The mean resistance rate against fosfomycin was 9.7% (range 7.1-11.3). No statistically significant difference was found between the three laboratories (P = 0.53). There was no significant increase in resistance rate during the study period. The mean resistance rate against fosfomycin was higher among extended-spectrum ß-lactamase (ESBL)-producing bacteria when compared with non-ESBL-producing strains (10.8% vs. 7.9%; P < 0.001). CONCLUSION: Uropathogens isolated from women affected by cystitis remained highly susceptible to fosfomycin. These findings confirm recommendations in international guidelines that advocate fosfomycin trometamol for empirical treatment of uncomplicated cystitis in women.

A Community Study of SARS-CoV-2 Detection by RT-PCR in Saliva: A Reliable and Effective Method.

Efficient, wide-scale testing for SARS-CoV-2 is crucial for monitoring the incidence of the infection in the community. The gold standard for COVID-19 diagnosis is the molecular analysis of epithelial secretions from the upper respiratory system captured by nasopharyngeal (NP) or oropharyngeal swabs. Given the ease of collection, saliva has been proposed as a possible substitute to support testing at the population level. Here, we used a novel saliva collection device designed to favour the safe and correct acquisition of the sample, as well as the processivity of the downstream molecular analysis. We tested 1003 nasopharyngeal swabs and paired saliva samples self-collected by individuals recruited at a public drive-through testing facility. An overall moderate concordance (68%) between the two tests was found, with evidence that neither system can diagnose the infection in 100% of the cases. While the two methods performed equally well in symptomatic individuals, their discordance was mainly restricted to samples from convalescent subjects. The saliva test was at least as effective as NP swabs in asymptomatic individuals recruited for contact tracing. Our study describes a testing strategy of self-collected saliva samples, which is reliable for wide-scale COVID-19 screening in the community and is particularly effective for contact tracing.

Rubella immunity status and the active offer of MMR/MMRV vaccination during pregnancy.

INTRODUCTION: The rubella test during pregnancy makes it possible to identify situations at risk of congenital rubella and those pregnant mothers who should be offered the MMR vaccine. MATERIALS AND METHODS: The Authors analysed test coverage and the immunity status of pregnant mothers between 2005 and 2017, using birth attendance certificates. RESULTS: Rubella test coverage on 61,437 pregnant mothers was 99.4%. The average proportion of susceptible subjects was 6.4%. Seroconversion was observed in 7 cases, with 1 confirmed case of congenital rubella. 32% of susceptible subjects were vaccinated, and adherence was seen to be influenced by the characteristics of the pregnant women and of the maternity unit. CONCLUSIONS: A current information flow including a number of healthcare services, is useful both for monitoring the maternity care pathway and for public health purposes.

Rhamnolipid coating reduces microbial biofilm formation on titanium implants: an in vitro study.

BACKGROUND: Peri-implant mucositis and peri-implantitis are biofilm-related diseases causing major concern in oral implantology, requiring complex anti-infective procedures or implant removal. Microbial biosurfactants emerged as new anti-biofilm agents for coating implantable devices preserving biocompatibility. This study aimed to assess the efficacy of rhamnolipid biosurfactant R89 (R89BS) to reduce Staphylococcus aureus and Staphylococcus epidermidis biofilm formation on titanium. METHODS: R89BS was physically adsorbed on titanium discs (TDs). Cytotoxicity of coated TDs was evaluated on normal lung fibroblasts (MRC5) using a lactate dehydrogenase assay. The ability of coated TDs to inhibit biofilm formation was evaluated by quantifying biofilm biomass and cell metabolic activity, at different time-points, with respect to uncoated controls. A qualitative analysis of sessile bacteria was also performed by scanning electron microscopy. RESULTS: R89BS-coated discs showed no cytotoxic effects. TDs coated with 4 mg/mL R89BS inhibited the biofilm biomass of S. aureus by 99%, 47% and 7% and of S. epidermidis by 54%, 29%, and 10% at 24, 48 and 72 h respectively. A significant reduction of the biofilm metabolic activity was also documented. The same coating applied on three commercial implant surfaces resulted in a biomass inhibition higher than 90% for S. aureus, and up to 78% for S. epidermidis at 24 h. CONCLUSIONS: R89BS-coating was effective in reducing Staphylococcus biofilm formation at the titanium implant surface. The anti-biofilm action can be obtained on several different commercially available implant surfaces, independently of their surface morphology.

The assessment of high sensitivity cardiac troponin in patients with COVID-19: A multicenter study.

BACKGROUND: Recent studies have shown that patients diagnosed with coronarivus disease 2019 (COVID-19) and also with previous cardiovascular diseases have a higher mortality due to worsening heart disease. At the same time, patients without previous cardiovascular disease may also have cardiac complications. The aim of this multicenter study was to assess high sensitivity cardiac troponin T (hs-cTnT) in patients with COVID-19 and to evaluate the incidence of myocardial injury. METHODS: In this multicenter study we enrolled 543 patients, 57.8% males, median age 63 years (range 18-99) from three selected hospitals: University Hospital Tor Vergata in Rome, Fondazione IRCCS Ca 'Granda Ospedale Maggiore Policlinico, in Milan, S Chiara Hospital in Trento. We measured hs-cTnT in all patients to assess myocardial injury and correlations with patient's age, symptoms and disease course. RESULTS: The data showed that, among the 543 patients studied, 257 patients (47.3%) had hs-cTnT values higher than the upper reference limit (URL) of 14 ng/L. Patients with high hs-cTnT had more frequently fever (p < 0.01) and respiratory symptoms (p < 0.01), compared to the group with hs-cTnT values below URL. The results showed also that patients with hs-cTnT above URL had a higher frequency of previous cardiovascular disease (p < 0.01) as well as of hypertension (p < 0.01). Instead, among 231 patients with no previous cardiovascular disease, 81 (31.5%) had hs-cTnT values above the URL. Finally. the majority of the patients with high hs-cTnT were admitted to the intensive care unit (p < 0.01). CONCLUSION: Our data suggest the assessment of high sensitivity cardiac troponin in patients with COVID-19 for early diagnosis of cardiac involvement.

Infectious Complications After Laser Vaporization of Urinary Stones During Retrograde Intrarenal Surgery Are Not Associated with Spreading of Bacteria into Irrigation Fluid but with Previous Use of Fluoroquinolones.

BACKGROUND: The pathogenesis of infectious complications after retrograde intrarenal surgery (RIRS) is not fully understood. OBJECTIVE: To evaluate spreading of bacteria into irrigation fluid and blood during RIRS for stone management and to correlate such spreading with infectious complications. DESIGN, SETTING, AND PARTICIPANTS: From January to December 2017, 38 patients who underwent RIRS for stones in two urological units were enrolled in this prospective, longitudinal cohort study. INTERVENTION: A urine culture was taken before surgery and antimicrobial prophylaxis was given in line with the European Association of Urology guidelines. Blood and irrigation fluid samples were collected at the start of the endoscopic procedure and every 30min during the procedure. All samples were microbiologically examined and findings were compared with clinical data. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSES: Symptomatic and asymptomatic urinary tract infectious complications were correlated with microbiological and clinical data, using Student t test or Mann-Whitney U test. RESULTS AND LIMITATIONS: Ten patients showed significant bacterial growth in irrigation fluid samples (seven Escherichia coli, two Klebsiella pneumoniae, and one Pseudomonas aeruginosa). Eight patients (21%) got febrile urinary tract infections during hospital stay: two had bacterial growth in the irrigation fluid (25%) and one also had bacteremia (12.5%). No correlation was found either between the bacterial growth in the irrigation fluid samples and the urine cultures that were taken before the procedure, or between the bacterial growth in the irrigation fluid samples and the development of postoperative infectious complications. Previous use of fluoroquinolones and a history of urinary tract infections were associated with infectious complications after RIRS. CONCLUSIONS: We demonstrated spreading of bacteria into the irrigation fluid during RIRS procedures, but this spreading was not associated with the development of infectious complications. Particular attention should be given to previous antibiotic treatment before administration of antimicrobial prophylaxis. PATIENT SUMMARY: Bacterial spreading into irrigation fluid is a common finding during retrograde intrarenal surgery, but it is not associated with infectious complications after the procedure. Particular attention should be given to previous antibiotic treatment before administration of antimicrobial prophylaxis.

Inhibitory Effects of Lipopeptides and Glycolipids on C. albicans-Staphylococcus spp. Dual-Species Biofilms.

Microbial biofilms strongly resist host immune responses and antimicrobial treatments and are frequently responsible for chronic infections in peri-implant tissues. Biosurfactants (BSs) have recently gained prominence as a new generation of anti-adhesive and antimicrobial agents with great biocompatibility and were recently suggested for coating implantable materials in order to improve their anti-biofilm properties. In this study, the anti-biofilm activity of lipopeptide AC7BS, rhamnolipid R89BS, and sophorolipid SL18 was evaluated against clinically relevant fungal/bacterial dual-species biofilms (Candida albicans, Staphylococcus aureus, Staphylococcus epidermidis) through quantitative and qualitative in vitro tests. C. albicans-S. aureus and C. albicans-S. epidermidis cultures were able to produce a dense biofilm on the surface of the polystyrene plates and on medical-grade silicone discs. All tested BSs demonstrated an effective inhibitory activity against dual-species biofilms formation in terms of total biomass, cell metabolic activity, microstructural architecture, and cell viability, up to 72 h on both these surfaces. In co-incubation conditions, in which BSs were tested in soluble form, rhamnolipid R89BS (0.05 mg/ml) was the most effective among the tested BSs against the formation of both dual-species biofilms, reducing on average 94 and 95% of biofilm biomass and metabolic activity at 72 h of incubation, respectively. Similarly, rhamnolipid R89BS silicone surface coating proved to be the most effective in inhibiting the formation of both dual-species biofilms, with average reductions of 93 and 90%, respectively. Scanning electron microscopy observations showed areas of treated surfaces that were free of microbial cells or in which thinner and less structured biofilms were present, compared to controls. The obtained results endorse the idea that coating of implant surfaces with BSs may be a promising strategy for the prevention of C. albicans-Staphylococcus spp. colonization on medical devices, and can potentially contribute to the reduction of the high economic efforts undertaken by healthcare systems for the treatment of these complex fungal-bacterial infections.

Dental Implants with Anti-Biofilm Properties: A Pilot Study for Developing a New Sericin-Based Coating.

AIM: several strategies have been tested in recent years to prevent bacterial colonization of dental implants. Sericin, one of the two main silk proteins, possesses relevant biological activities and also literature reports about its potential antibacterial properties, but results are discordant and not yet definitive. The aim of this study was to evaluate the effectiveness of different experimental protocols in order to obtain a sericin-based coating on medical grade titanium (Ti) able to reduce microbial adhesion to the dental implant surface. MATERIALS AND METHODS: different strategies for covalent bonding of sericin to Ti were pursued throughout a multi-step procedure on Ti-6Al-4V disks. The surface of grade 5 Ti was initially immersed in NaOH solution to obtain the exposure of functional -OH groups. Two different silanization strategies were then tested using aminopropyltriethoxysilane (APTES). Eventually, the bonding between silanized Ti-6Al-4V and sericin was obtained with two different crosslinking processes: glutaraldehyde (GLU) or carbodiimide/N-Hydroxy-succinimide (EDC/NHS). Micro-morphological and compositional analyses were performed on the samples at each intermediate step to assess the most effective coating strategy able to optimize the silanization and bioconjugation processes. Microbiological tests on the coated Ti-6Al-4V disks were conducted in vitro using a standard biofilm producer strain of Staphylococcus aureus (ATCC 6538) to quantify the inhibition of microbial biofilm formation (anti-biofilm efficacy) at 24 hours. RESULTS: both silanization techniques resulted in a significant increase of silicon (Si) on the Ti-6Al-4V surfaces etched with NaOH. Differences were found between GLU and EDC/NHS bioconjugation strategies in terms of composition, surface micro-morphology and anti-biofilm efficacy. Ti-6Al-4V samples coated with GLU-bound sericin after silanization obtained via vapor phase deposition proved that this technique is the most convenient and effective coating strategy, resulting in a bacterial inhibition of about 53% in respect to the uncoated Ti-6Al-4V disks. CONCLUSIONS: The coating with glutaraldehyde-bound sericin after silanization in the vapor phase showed promising bacterial inhibition values with a significant reduction of S. aureus biofilm. Further studies including higher number of replicates and more peri-implant-relevant microorganisms are needed to evaluate the applicability of this experimental protocol to dental implants.

Microscopic Observation of Proliferative Membranes in Fibrocontractive Retinal Disorders.

Proliferative membranes of fibrocontractive retinal disorders are extensively studied from the morphological and evolutive point of view. Despite this, little is known of their cellular composition. In this study, the authors investigated the morphological characteristics and cell composition of various types of surgically excised proliferative membranes and internal limiting membranes (ILMs), in order to provide new data supporting or challenging the pathogenic theories proposed until now. Sixty-nine specimens from 64 eyes of 64 consecutive patients were collected at surgery and subjected to a multilevel analysis by means of optical and electron microscopy. Membrane samples were semiquantitatively evaluated for the amount and distribution of cell nuclei and pigment. Immunohistochemical staining was performed with antibodies to alpha smooth muscle actin and CD68. Data were analyzed after grouping according to the following tissue types: ILM (20 specimens), epiretinal membrane (ERM) (22 specimens), ILM + ERM (20 specimens), and proliferative vitreoretinopathy (PVR) (7 specimens). The cell components found in the ERM specimens, like myofibroblasts, macrophages, and polymorphonuclear cells, were recognized as the expression of cell migration and differentiation that induced an inflammatory process and a fibroproliferative repair process. The detection of pigments in specific types of ERM, like those associated with lamellar macular hole (LMH) or secondary to retinal detachment (RD), diabetes, and PVR, suggested that retinal pigment epithelium (RPE) cells may have a role in the development of these vitreoretinal disorders. The reduction of the ERM cellularity with the patient's age supports the hypothesis that ERM evolves in time up to a fibrous tissue formation.

Xyloglucan, hibiscus and propolis to reduce symptoms and antibiotics use in recurrent UTIs: a prospective study.

Aim: To evaluate the efficacy of a medical device containing xyloglucan, hibiscus and propolis in the management of recurrent urinary tract infections (rUTIs). Patients & methods: Sixty-one women affected by rUTIs received this medical device, one capsule a day for 15 days (one cycle every month, for 6 months), in an observational, prospective study. Clinical and microbiological evaluations were performed at baseline and 1, 3 and 6 months from enrolment. Results: At first follow-up, 41 reported a clinical improvement and a return to their clinical status before UTI, while 47 and 51 did so at the second and third follow-up evaluations. A statistically significant clinical improvement was reported at each follow-up visit (quality of life [QoL] 94.2 vs 98.6; QoL 94.1 vs 98.7; QoL 94.2 vs 99.1; p < 0.001). A statistically significant reduction in antibiotic use was reported. Conclusion: This medical device is able to improve quality of life in women with rUTIs, reduce recurrences and antibiotic use.

Role of increasing leukocyturia for detecting the transition from asymptomatic bacteriuria to symptomatic infection in women with recurrent urinary tract infections: A new tool for improving antibiotic stewardship.

OBJECTIVES: To evaluate the importance of leukocyturia in detecting the transition from asymptomatic bacteriuria to symptomatic infection in women with recurrent urinary tract infections. METHODS: In this cross-sectional study, we evaluated all women with recurrent urinary tract infection and asymptomatic bacteriuria who had been enrolled in two previous studies. Data from urological visits, urine analyses and microbiological evaluations were collected. Patients were divided into two groups: patients with symptomatic recurrence (group A) and patients without recurrence (group B), with a mean follow-up period of 38.8 months. Data on leukocyturia and clinical data were compared. Logistic regression analyses were carried out and areas under the receiver operating characteristic curves were calculated. RESULTS: A total of 301 women with symptomatic urinary tract infection were included in group A, whereas 249 women without clinical infection were included in group B. Group A showed a higher level of leukocytes in the urinary analysis taken at the moment of recurrence when compared with the baseline value (mean leukocytes per high power field 54 ± 5 vs 19 ± 6 at baseline; P < 0.0001). When an increase of leukocytes/mm3 of >150% from baseline was used for logistic regression, the area under the receiver operating characteristic of the model was 0.82 (95% CI 0.78-0.94; P = 0.01). An increase of leukocytes/mm3 of >150% from baseline had a sensitivity of 90.1% and a specificity of 91.2% for symptomatic urinary tract infection. CONCLUSIONS: This study shows that an increase of leukocyturia of >150% from baseline has a predictive role for the transition from asymptomatic bacteriuria to symptomatic urinary tract infection in women with recurrent urinary tract infections.

Prostate calcifications: A case series supporting the microbial biofilm theory.

Purpose: Prostate calcifications are a common finding during transrectal prostate ultrasound in both healthy subjects and patients, but their etiopathogenesis and clinical significance are not fully understood. We aimed to establish a new methodology for evaluating the role of microbial biofilms in the genesis of prostate calcifications. Materials and Methods: Ten consecutive patients who had undergone radical prostatectomy were enrolled in this study. All of the patients presented with prostate calcifications during transrectal ultrasound evaluation before surgery and underwent Meares-Stamey tests and clinical evaluation with the National Institutes of Health Chronic Prostatitis Symptom Index and the International Prostate Symptom Score. At the time of radical prostatectomy, the prostate specimen, after removal, was analyzed with ultrasonography under sterile conditions in the operating room. Core biopsy specimens were taken from the site of prostate calcification and subjected to ultrastructural and microbiological analysis. Results: The results of the Meares-Stamey test showed only 1 of 10 patients (10%) with positive cultures for Escherichia coli. Two of five patients (40%) had positive cultures from prostate biopsy specimens. Enterococcus faecalis, Enterococcus raffinosus, and Citrobacter freundii were isolated. Ultrastructural analysis of the prostate biopsy specimens showed prostate calcifications in 6 of 10 patients (60%), and a structured microbial biofilm in 1 patient who had positive cultures for E. faecalis and E. raffinosus. Conclusions: Although the findings are supported by a low number of patients, this study highlights the validity of the proposed methodology for investigating the role of bacterial biofilms in the genesis of prostate calcification.

The Efficacy of Umbelliferone, Arbutin, and N-Acetylcysteine to Prevent Microbial Colonization and Biofilm Development on Urinary Catheter Surface: Results from a Preliminary Study.

We evaluated, in a preliminary study, the efficacy of umbelliferone, arbutin, and N-acetylcysteine to inhibit biofilm formation on urinary catheter. We used 20 urinary catheters: 5 catheters were incubated with Enterococcus faecalis (control group); 5 catheters were incubated with E. faecalis in presence of umbelliferone (150 mg), arbutin (60 mg), and N-acetylcysteine (150 mg) (group 1); 5 catheters were incubated with E. faecalis in presence of umbelliferone (150 mg), arbutin (60 mg), and N-acetylcysteine (400 mg) (group 2); and 5 catheters were incubated with E. faecalis in presence of umbelliferone (300 mg), arbutin (60 mg), and N-acetylcysteine (150 mg) (group 3). After 72 hours, planktonic microbial growth and microorganisms on catheter surface were assessed. In the control group, we found a planktonic load of ≥10(5) CFU/mL in the inoculation medium and retrieved 3.69 × 10(6) CFU/cm from the sessile cells adherent to the catheter surface. A significantly lower amount in planktonic (p < 0.001) and sessile (p = 0.004) bacterial load was found in group 3, showing <100 CFU/mL and 0.12 × 10(6) CFU/cm in the incubation medium and on the catheter surface, respectively. In groups 1 and 2, 1.67 × 10(6) CFU/cm and 1.77 × 10(6) CFU/cm were found on catheter surface. Our results document that umbelliferone, arbutin, and N-acetylcysteine are able to reduce E. faecalis biofilm development on the surface of urinary catheters.

Asymptomatic Bacteriuria in Clinical Urological Practice: Preoperative Control of Bacteriuria and Management of Recurrent UTI.

Asymptomatic bacteriuria (ABU) is a common clinical condition that often leads to unnecessary antimicrobial use. The reduction of antibiotic overuse for ABU is consequently an important issue for antimicrobial stewardship and to reduce the emergence of multidrug resistant strains. There are two issues in everyday urological practice that require special attention: the role of ABU in pre-operative prophylaxis and in women affected by recurrent urinary tract infections (rUTIs). Nowadays, this is the time to think over our practice and change our way of thinking. Here, we aimed to summarize the current literature knowledge in terms of ABU management in patients undergoing urological surgery and in patients with rUTIs. In the last years, the approach to patient with ABU has changed totally. Prior to all surgical procedures that do not enter the urinary tract, ABU is generally not considered as a risk factor, and screening and treatment are not considered necessary. On the other hand, in the case of all procedures entering the urinary tract, ABU should be treated in line with the results of a urine culture obtained before the procedure. In patients affected by rUTIs, ABU can even have a protective role in preventing symptomatic recurrence, particularly when Enterococcus faecalis (E. faecalis) has been isolated.

Asymptomatic bacteriuria treatment is associated with a higher prevalence of antibiotic resistant strains in women with urinary tract infections.

BACKGROUND: Women suffering from recurrent urinary tract infections (rUTIs) are routinely treated for asymptomatic bacteriuria (AB), but the consequences of this procedure on antibiotic resistance are not fully known. The aim of this study was to evaluate the impact of AB treatment on antibiotic resistance among women with rUTIs. METHODS: The study population consisted of 2 groups of women who had previously been enrolled in a randomized clinical trial: group A was not treated, and group B was treated. All women were scheduled for follow-up visits every 6 months, or more frequently if symptoms arose. Microbiological evaluation was performed only in symptomatic women. All women were followed up for a mean of 38.8 months to analyze data from urine cultures and antibiograms. RESULTS: The previous study population consisted of 673 women, but 123 did not attend the entire follow-up period. For the final analysis, 257 of the remaining 550 patients were assigned to group A, and 293 to group B. At the end of follow-up, the difference in recurrence rates was statistically significant (P < .001): 97 (37.7%) in group A versus 204 (69.6%) in group B. Isolated Escherichia coli from group B showed higher resistance to amoxicillin-clavulanic acid (P = .03), trimethoprim-sulfamethoxazole (P = .01), and ciprofloxacin (P = .03) than that from group A. CONCLUSIONS: This study shows that AB treatment is associated with a higher occurrence of antibiotic-resistant bacteria, indicating that AB treatment in women with rUTIs is potentially dangerous.

Solidago, orthosiphon, birch and cranberry extracts can decrease microbial colonization and biofilm development in indwelling urinary catheter: a microbiologic and ultrastructural pilot study.

PURPOSE: Plants extracts are used in urology to manage urinary tract infections. We aimed to evaluate the efficacy of a preparation with solidago, orthosiphon, birch and cranberry extracts (CISTIMEV PLUS(®)) in reducing microbial colonization and biofilm development in patients with indwelling urinary catheters. METHODS: All consecutive outpatients attending our department between January and June 2010 for the substitution of indwelling catheters were considered for this single-blinded, randomized and controlled pilot study to test superiority of the preventative management (CISTIMEV PLUS(®), 1 tablet daily for 30 days) in respect to no treatment. A sample size of 10-40 participants per group was considered adequate. All patients underwent urine culture the same day of the catheter substitution and were then randomized into test group (n = 48) and control group (n = 35). Ultrastructural analysis was also performed. After 30 days, the catheter was replaced and the analysis repeated. The primary outcome was the rate of positive urinary culture at the end of the entire study period. RESULTS: Ten patients abandoned the study. At 30 days, according to per-protocol analysis, the groups statistically differed regarding the rate of positive urine cultures: test group 10/43 and control group 16/30 (p = 0.013) (-30.1 % [95 % CI -51.94 to -8.21]). The most common isolated bacteria were Escherichia coli and Enterococcus faecalis. CONCLUSIONS: The use of solidago, orthosiphon, birch and cranberry extracts resulted in a significant reduction of microbial colonization in patients with indwelling urinary catheters. Larger clinical trials are needed to demonstrate that the effects here reported are sufficient to reduce symptomatic catheter-associated urinary tract infections.

Quality Indicators in Laboratory Medicine: from theory to practice. Preliminary data from the IFCC Working Group Project "Laboratory Errors and Patient Safety".

BACKGROUND: The adoption of Quality Indicators (QIs) has prompted the development of tools to measure and evaluate the quality and effectiveness of laboratory testing, first in the hospital setting and subsequently in ambulatory and other care settings. While Laboratory Medicine has an important role in the delivery of high-quality care, no consensus exists as yet on the use of QIs focussing on all steps of the laboratory total testing process (TTP), and further research in this area is required. METHODS: In order to reduce errors in laboratory testing, the IFCC Working Group on "Laboratory Errors and Patient Safety" (WG-LEPS) developed a series of Quality Indicators, specifically designed for clinical laboratories. In the first phase of the project, specific QIs for key processes of the TTP were identified, including all the pre-, intra- and post-analytic steps. The overall aim of the project is to create a common reporting system for clinical laboratories based on standardized data collection, and to define state-of-the-art and Quality Specifications (QSs) for each QI independent of: a) the size of organization and type of activities; b) the complexity of processes undertaken; and c) different degree of knowledge and ability of the staff. The aim of the present paper is to report the results collected from participating laboratories from February 2008 to December 2009 and to identify preliminary QSs. RESULTS AND CONCLUSIONS: The results demonstrate that a Model of Quality Indicators managed as an External Quality Assurance Program can serve as a tool to monitor and control the pre-, intra- and post-analytical activities. It might also allow clinical laboratories to identify risks that lead to errors resulting in patient harm: identification and design of practices that eliminate medical errors; the sharing of information and education of clinical and laboratory teams on practices that reduce or prevent errors; the monitoring and evaluation of improvement activities.

Sterility and microbiological assessment of reused single-use cardiac electrophysiology catheters.

OBJECTIVE: To assess the performance and limitations of a reprocessing protocol for nonlumen electrophysiology catheters by testing the sterility of reprocessed devices and defining the maximum number of reprocessing cycles sustainable by the device in hygienically safe conditions. DESIGN: Simulated use, reprocessing, and testing of the catheters. SETTING: Microbiology and virology department of a public health diagnostic laboratory. INTERVENTIONS: Seventy-three catheters were collected after clinical use on patients. The first group of devices was tested for sterility after 1 cycle of reprocessing. By the repetition of simulated use (blood inoculated with bacteria) and reprocessing (decontamination, cleaning, and hydrogen peroxide gas plasma sterilization), we obtained 39 sample devices reprocessed 2 times, 26 reprocessed 3 times, 28 reprocessed 4 times, 36 reprocessed 5 times, and 22 reprocessed 6 times. Devices were cultured for 28 days in trypticase soy broth. RESULTS: We tested 208 catheters with 6 cycles of reprocessing and 4 inoculated bacteria species. No devices tested positive for the inoculated strains until the fourth cycle of reprocessing. One of 35 catheters showed the growth of the inoculated strain Bacillus subtilis after 5 cycles of reprocessing, and 1 of 22 catheters showed growth of this organism 6 cycles. After the second reprocessing, 7 of 36 devices showed growth of gram-negative bacteria other than the strain inoculated. CONCLUSIONS: Reprocessing according to the reprocessing protocol was insufficient to guarantee device sterility after 5 reuses. Cleaning with enzymatic solution revealed good cleaning properties with efficient bioburden reduction. Storage intervals of longer than 24 hours during reprocessing should be avoided to limit contamination or bacterial overgrowth. Technical considerations suggest the introduction of reprocessing procedures only in hospitals with considerable workloads.

Efficiency in endotoxin removal by a reprocessing protocol for electrophysiology catheters based on hydrogen peroxide plasma sterilization.

Electrophysiology and ablation cardiac catheters, which come in contact with blood during clinical use, are required to be non-pyrogenic (<20 endotoxin units (EU)/device). This study aimed to quantify the residual endotoxin load in reprocessed devices as a mandatory step to guarantee safe reuse. We monitored the pyrogenic status of the device (n=61) in three fundamental steps of the reprocessing protocol: after clinical use, after decontamination-cleaning treatments and after complete reprocessing, including sterilization by hydrogen peroxide gas plasma. Finally, a depyrogenation test was produced for evaluating the depyrogenation efficiency of the sole hydrogen peroxide sterilization treatment. Results showed that standard clinical use did not represent a source for endotoxin contamination, while the use of tap water and manual cleaning processing could increase the pyrogenic load in a significant way. The introduction of the sterilization by hydrogen peroxide gas plasma resulted in effective reduction of the endotoxin contamination and in safe reprocessing of 15 of 15 clinically used catheters. In addition, tests conducted on in vitro spiked catheters showed that initial pyrogenic loads of 40, 80, 200EU/device were reduced to less than 11EU/device. Depyrogenation testing demonstrated efficiency in endotoxin reduction of more than 62 times (1.8log). These results show the determining role of hydrogen peroxide gas-plasma sterilization in the reduction of pyrogenic load on medical devices. Considering actual hygienic requirements at single-use device reprocessing, hydrogen peroxide gas-plasma sterilization can be considered as an efficient treatment at non-lumen cardiac electrophysiology catheter reprocessing.

Diagnostic accuracy of ELISA methods as an alternative screening test to indirect immunofluorescence for the detection of antinuclear antibodies. Evaluation of five commercial kits.

Detection of antinuclear antibodies (ANA) is a fundamental laboratory test for diagnosing systemic autoimmune diseases. Currently, the method of choice is indirect immunofluorescence (IIF) on a HEp-2 cell substrate. The goal of this study was to evaluate the diagnostic accuracy of five commercially available enzyme immunoassay (EIA) kits for ANA detection and to verify the possibility of using them as an alternative to the IIF method. The study involved 1513 patients, 315 of whom were diagnosed with a systemic autoimmune disease and 1198 in whom an autoimmune disorder was excluded. For all sera, ANA detection was performed via IIF and with five different EIA kits. The results were evaluated in relation to clinical diagnosis and the presence of possible specific autoantibodies (anti-ENA or anti-dsDNA); lastly, they were compared with the results obtained using ANA-IIF as the method of reference. The positive rate of the ANA-IIF test in subjects with systemic autoimmune diseases was 92%, whereas in the five ANA-EIA kits there was broad diversity in terms of response, with positive rates ranging from 74 to 94%. All the EIA kits correctly detected the presence of antibodies (anti-dsDNA, anti-RNP, anti-Ro/SSA) responsible for homogeneous and speckled fluorescence pattern, but at the same time they showed substantial inaccuracy with the nucleolar pattern, with a mean sensitivity of approximately 50% in this case. Instead, there was a large kit-to-kit difference in terms of identification of anti-Scl70 and centromere patterns, for which sensitivities ranged between 45 and 91%, and between 49 and 100%, respectively. The results of the study demonstrate that the commercially available ANA-EIA kits show different levels of sensitivity and specificity. Some of them have a diagnostic accuracy that is comparable and, in some cases, even higher than the IIF method. Consequently, these could be used as an alternative screening test to IIE. However, others do not ensure acceptable results. Therefore, careful evaluation of the various kits on the market is advisable before including any of these methods in the clinical and diagnostic testing.