Biomarkers to identify Mycobacterium tuberculosis infection among borderline QuantiFERON results.

BACKGROUND: Screening for tuberculosis (TB) infection often includes QuantiFERON-TB Gold Plus (QFT) testing. Previous studies showed that two-thirds of patients with negative QFT results just below the cut-off, so-called borderline test results, nevertheless had other evidence of TB infection. This study aimed to identify a biomarker profile by which borderline QFT results due to TB infection can be distinguished from random test variation. METHODS: QFT supernatants of patients with a borderline (≥0.15 and <0.35 IU·mL-1), low-negative (<0.15 IU·mL-1) or positive (≥0.35 IU·mL-1) QFT result were collected in three hospitals. Bead-based multiplex assays were used to analyse 48 different cytokines, chemokines and growth factors. A prediction model was derived using LASSO regression and applied further to discriminate QFT-positive Mycobacterium tuberculosis-infected patients from borderline QFT patients and QFT-negative patients RESULTS: QFT samples of 195 patients were collected and analysed. Global testing revealed that the levels of 10 kDa interferon (IFN)-γ-induced protein (IP-10/CXCL10), monokine induced by IFN-γ (MIG/CXCL9) and interleukin-1 receptor antagonist in the antigen-stimulated tubes were each significantly higher in patients with a positive QFT result compared with low-negative QFT individuals (p<0.001). A prediction model based on IP-10 and MIG proved highly accurate in discriminating patients with a positive QFT (TB infection) from uninfected individuals with a low-negative QFT (sensitivity 1.00 (95% CI 0.79-1.00) and specificity 0.95 (95% CI 0.74-1.00)). This same model predicted TB infection in 68% of 87 patients with a borderline QFT result. CONCLUSIONS: This study was able to classify borderline QFT results as likely infection-related or random. These findings support additional laboratory testing for either IP-10 or MIG following a borderline QFT result for individuals at increased risk of reactivation TB.

Evaluation of the Liat Cdiff Assay for Direct Detection of Clostridioides difficile Toxin Genes within 20 Minutes.

Clostridioides difficile is the main causative agent of antibiotic-associated diarrhea. Prompt diagnosis is required for initiation of timely infection control measures and appropriate adjustment of antibiotic treatment. The cobas Cdiff assay for use on the cobas Liat system enables a diagnostic result in 20 minutes. A total of 252 prospective (n = 150) and retrospective (n = 102) stool specimens from The Netherlands, France, and Switzerland were tested on the cobas Cdiff assay using the Xpert C. difficile assay as a reference method. The overall positive and negative percent agreement (PPA and NPA, respectively) of the cobas Cdiff assay compared with the Xpert C. difficile assay was 98.0% (100/102; 95% confidence interval [CI], 93.1% to 99.5%) and 94.0% (141/150; 95% CI, 89.0% to 96.8%), respectively. When comparing the PPAs of cobas Cdiff and Xpert C. difficile with culture, the results were 91.7% (55/60; 95% CI, 81.9% to 96.4%) and 85.0% (51/60; 95% CI, 73.9% to 91.9%), respectively. The difference was not statistically significant. The cobas Cdiff assay offers a very rapid alternative for diagnosing C. difficile infection. The 20-minute turnaround time provides the potential for point-of-care testing so that adequate infection control measures can be initiated promptly.

Prevention of Surgical Site Infections: Decontamination With Mupirocin Based on Preoperative Screening for Staphylococcus aureus Carriers or Universal Decontamination?

Perioperative decolonization of Staphylococcus aureus nasal carriers with mupirocin together with chlorhexidine body washing reduces the incidence of S. aureus surgical site infection. A targeted strategy, applied in S. aureus carriers only, is costly, and implementation may reduce effectiveness. Universal decolonization is more cost-effective but increases exposure of noncarriers to mupirocin and the risk of resistance to mupirocin in staphylococci. High-level mupirocin resistance in S. aureus can emerge through horizontal gene transfer originating from coagulase-negative staphylococci (CoNS) and through clonal transmission. The current evidence on the occurrence of high-level mupirocin resistance in S. aureus and CoNS, in combination with the results of mathematical modeling, strongly suggests that the increased selection of high-level mupirocin resistance in CoNS does not constitute an important risk for high-level mupirocin resistance in S. aureus. Compared with a targeted strategy, universal decolonization seems associated with an equally low risk of mupirocin resistance in S. aureus.

Acquisition of high-level mupirocin resistance in CoNS following nasal decolonization with mupirocin.

OBJECTIVES: The association between mupirocin use and plasmid-based high-level resistance development mediated through mupA in CoNS has not been quantified. We determined acquisition of mupirocin resistance in Staphylococcus aureus and CoNS in surgery patients treated peri-operatively with mupirocin. PATIENTS AND METHODS: Patients admitted for surgery were treated with nasal mupirocin ointment and chlorhexidine soap for 5 days, irrespective of S. aureus carrier status. Nasal swabs were obtained before decolonization (T1) and 4 days after surgery (T2) and were inoculated onto agars containing 8 mg/L mupirocin. Staphylococci were identified by MALDI-TOF MS and mupirocin resistance was confirmed by Etest. RESULTS: Among 1578 surgical patients, 936 (59%) had nasal swabs obtained at T1 and T2; 192 (21%) patients carried mupirocin-resistant CoNS at T1 and 406 (43%) at T2 (P<0.001). Of 744 patients not colonized at T1, 277 acquired resistance (37%), corresponding to an acquisition rate of 7.4/100 patient days at risk. In all, 588 (97%) of 607 mupirocin-resistant CoNS had an MIC >256 mg/L (high level) and 381 of 383 (99.5%) were mupA positive. No acquisition of mupirocin resistance was observed in S. aureus. CONCLUSIONS: Acquisition of mupirocin resistance following decolonization was widespread in CoNS and absent in S. aureus. As almost all isolates harboured the mupA gene, monitoring resistance development in S. aureus when decolonization strategies containing mupirocin are used is recommended.

Transmissibility of livestock-associated methicillin-resistant Staphylococcus aureus.

Previous findings have suggested that the nosocomial transmission capacity of livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is lower than that of other MRSA genotypes. We therefore performed a 6-month (June 1-November 30, 2011) nationwide study to quantify the single-admission reproduction number, RA, for LA-MRSA in 62 hospitals in the Netherlands and to compare this transmission capacity to previous estimates. We used spa typing for genotyping. Quantification of RA was based on a mathematical model incorporating outbreak sizes, detection rates, and length of hospital stay. There were 141 index cases, 40 (28%) of which were LA-MRSA. Contact screening of 2,101 patients and 7,260 health care workers identified 18 outbreaks (2 LA-MRSA) and 47 secondary cases (3 LA-MRSA). RA values indicated that transmissibility of LA-MRSA is 4.4 times lower than that of other MRSA (not associated with livestock).

[Urinary tract infection caused by Candida while using an SGLT-2 inhibitor]. / Urineweginfectie door Candida bij gebruik SGLT2-remmer.

BACKGROUND: Sodium glucose transporter 2 (SGLT-2) inhibitors are a relatively new group of antidiabetic drugs. The renal glucose reabsorption is blocked resulting in higher glucose levels in the urine (glucosuria). Recently studies are being conducted into the medications nephrological and cardiovascular potential. As a result, we may expect that SGLT-2 inhibitors will be more and more frequently prescribed. Thus, physicians of any specialty may come into contact with patients that are using this drug. CASE DESCRIPTION: We describe a case of a male patient who developed a urinary tract infection with Candida glabrata while using a SGLT-2 inhibitor. After discontinuing the SGLT-2 inhibitor, the infection subsided. CONCLUSION: Urinary tract infections from Candida are rarely seen in healthy individuals. Glucosuria is a known risk factor for fungal genital infections. More research is needed to determine whether SGLT-2 inhibitors increase the risk of fungal urinary tract infections.

Semiautomated surveillance of deep surgical site infections after colorectal surgeries: A multicenter external validation of two surveillance algorithms.

OBJECTIVE: Automated surveillance methods increasingly replace or support conventional (manual) surveillance; the latter is labor intensive and vulnerable to subjective interpretation. We sought to validate 2 previously developed semiautomated surveillance algorithms to identify deep surgical site infections (SSIs) in patients undergoing colorectal surgeries in Dutch hospitals. DESIGN: Multicenter retrospective cohort study. METHODS: From 4 hospitals, we selected colorectal surgery patients between 2018 and 2019 based on procedure codes, and we extracted routine care data from electronic health records. Per hospital, a classification model and a regression model were applied independently to classify patients into low- or high probability of having developed deep SSI. High-probability patients need manual SSI confirmation; low-probability records are classified as no deep SSI. Sensitivity, positive predictive value (PPV), and workload reduction were calculated compared to conventional surveillance. RESULTS: In total, 672 colorectal surgery patients were included, of whom 28 (4.1%) developed deep SSI. Both surveillance models achieved good performance. After adaptation to clinical practice, the classification model had 100% sensitivity and PPV ranged from 11.1% to 45.8% between hospitals. The regression model had 100% sensitivity and 9.0%-14.9% PPV. With both models, <25% of records needed review to confirm SSI. The regression model requires more complex data management skills, partly due to incomplete data. CONCLUSIONS: In this independent external validation, both surveillance models performed well. The classification model is preferred above the regression model because of source-data availability and less complex data-management requirements. The next step is implementation in infection prevention practices and workflow processes.

Emergence of high-level mupirocin resistance in coagulase-negative staphylococci associated with increased short-term mupirocin use.

In our hospital, mupirocin has increasingly been used for peri-operative decolonization of Staphylococcus aureus. The target for mupirocin is isoleucyl tRNA synthetase (ileS). High-level resistance to mupirocin is conferred by acquisition of plasmids expressing a distinct ileS gene (ileS2). Here we evaluated the longitudinal trends in high-level mupirocin resistance in coagulase-negative staphylococci (CoNS) and linked this to the presence of ileS2 genes and mupirocin use. We assessed mupirocin resistance in CoNS bloodstream isolates from 2006 to 2011 tested by Phoenix automated testing (PAT). We evaluated the reliability of PAT results using Etest. PAT species determination was confirmed by MALDI-TOF (matrix-assisted laser desorption ionization-time of flight) mass spectrometry. We investigated the presence of ileS2 in the first 100 consecutive CoNS bloodstream isolates of each year using RT-PCR. Mupirocin use increased from 3.6 kg/year in 2006 to 13.3 kg/year in 2010 and correlated with the increase in the percentage of CoNS isolates carrying ileS2 (8% in 2006 to 22% in 2011; Spearman's rho, 0.137; P = 0.01). The sensitivity and specificity of PAT for detecting high-level mupirocin resistance were 0.97 and 0.97, respectively. ileS2 was detected in 81 of 82 phenotypically highly mupirocin-resistant strains and associated with resistance to ciprofloxacin, erythromycin, and clindamycin. In conclusion, we found a rapid increase in high-level resistance to mupirocin and resistance to other antibiotics in CoNS associated with an increase in mupirocin use. The associated resistance to other antibiotics may result in a reduction of oral antibiotic options for prolonged treatment of prosthetic infections with CoNS.

Nosocomial transmission of community-associated methicillin-resistant Staphylococcus aureus in Danish Hospitals.

OBJECTIVES: The emergence of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has changed the epidemiology of MRSA infections worldwide. In contrast to hospital-associated MRSA (HA-MRSA), CA-MRSA more frequently affects healthy individuals, both with and without recent healthcare exposure. Despite obvious epidemiological differences, it is unknown whether differences in nosocomial transmissibility exist. We have, therefore, quantified the transmissibility, expressed by the single admission reproduction number (R(A)), of CA-MRSA and HA-MRSA in hospital settings in Denmark. METHODS: MRSA index cases and secondary cases were investigated in four hospitals in the Copenhagen area. Index cases were defined as non-isolated, non-screened patients with MRSA, and secondary cases were defined as persons carrying MRSA isolates-identical to that of the corresponding index-as identified through contact screening. CA-MRSA and HA-MRSA were categorized upon genotyping [CA-MRSA: t008-ST8, PVL+; t019-ST30, PVL+; t127-ST1, PVL+; t044-ST80, PVL+; and their related spa types; and HA-MRSA: all other (where ST stands for sequence type and PVL stands for Panton-Valentine leucocidin)]. A mathematical model was applied to determine the genotype-specific transmission rate (i.e. R(A)) of CA-MRSA and HA-MRSA strains. RESULTS: During the 7 year study period there were 117 MRSA index cases with subsequent post-contact screening (of 1108 patients and healthcare workers), revealing 22 outbreaks with a total of 52 secondary patients. R(A) values were 0.07 (95% CI 0.00-0.28) and 0.65 (95% CI 0.48-0.84) for CA-MRSA and HA-MRSA, respectively. CONCLUSIONS: In four Danish hospitals the nosocomial transmission rate of CA-MRSA was 9.3 times lower than that of HA-MRSA.

Livestock-associated methicillin-resistant Staphylococcus aureus epidemiology, genetic diversity, and clinical characteristics in an urban region.

Objectives: While Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA), defined as CC398, is a well-known pathogen among those working with livestock, there are indications that LA-MRSA prevalence among the general population is increasing. However, the clinical impact in urban areas remains unknown. The aim of this study was to assess the genetic epidemiology and clinical characteristics of LA-MRSA in an urban area with a limited livestock population. Methods: In this retrospective study, we evaluated LA-MRSA strains that were collected between 2014 and 2018 from patients who received clinical care in a single urban area in Netherlands. Patient files were assessed for livestock exposure data, clinical findings, and contact tracing information. Next-generation sequencing (NGS) analysis in combination with wgMLST was conducted to assess genetic diversity and relatedness and to detect virulence and resistance genes. Results: LA-MRSA strains were cultured from 81 patients, comprising 12% of all the MRSA strains found in seven study laboratories between 2014 and 2018. No livestock link was found in 76% of patients (n = 61), and 28% of patients (n = 23) had an infection, mostly of the skin or soft tissue. Contact tracing had been initiated in 14 cases, leading to the identification of two hospital transmissions: a cluster of 9 cases and one of 2 cases. NGS data were available for 91% (n = 75) of the patients. wgMLST confirmed the clusters detected via contact tracing (n = 2) and identified 5 additional clusters without a known epidemiological link. Relevant resistance and virulence findings included the PVL virulence gene (3 isolates) and tetracycline resistance (79 isolates). Conclusion: LA-MRSA may cause a relevant burden of disease in urban areas. Surprisingly, most infections in the present study occurred in the absence of a livestock link, suggesting inter-human transmission. These findings and the presence of PVL and other immune evasive complex virulence genes warrant future surveillance and preventative measures.

Reliability and validity of multicentre surveillance of surgical site infections after colorectal surgery.

BACKGROUND: Surveillance is the cornerstone of surgical site infection prevention programs. The validity of the data collection and awareness of vulnerability to inter-rater variation is crucial for correct interpretation and use of surveillance data. The aim of this study was to investigate the reliability and validity of surgical site infection (SSI) surveillance after colorectal surgery in the Netherlands. METHODS: In this multicentre prospective observational study, seven Dutch hospitals performed SSI surveillance after colorectal surgeries performed in 2018 and/or 2019. When executing the surveillance, a local case assessment was performed to calculate the overall percentage agreement between raters within hospitals. Additionally, two case-vignette assessments were performed to estimate intra-rater and inter-rater reliability by calculating a weighted Cohen's Kappa and Fleiss' Kappa coefficient. To estimate the validity, answers of the two case-vignettes questionnaires were compared with the answers of an external medical panel. RESULTS: 1111 colorectal surgeries were included in this study with an overall SSI incidence of 8.8% (n = 98). From the local case assessment it was estimated that the overall percent agreement between raters within a hospital was good (mean 95%, range 90-100%). The Cohen's Kappa estimated for the intra-rater reliability of case-vignette review varied from 0.73 to 1.00, indicating substantial to perfect agreement. The inter-rater reliability within hospitals showed more variation, with Kappa estimates ranging between 0.61 and 0.94. In total, 87.9% of the answers given by the raters were in accordance with the medical panel. CONCLUSIONS: This study showed that raters were consistent in their SSI-ascertainment (good reliability), but improvements can be made regarding the accuracy (moderate validity). Accuracy of surveillance may be improved by providing regular training, adapting definitions to reduce subjectivity, and by supporting surveillance through automation.

High prevalence and resistance rates of Mycoplasma genitalium among patients visiting two sexually transmitted infection clinics in the Netherlands.

Mycoplasma genitalium is a well-known cause of urethritis in men and has been associated with cervicitis, pelvic inflammatory disease, and adverse obstetric outcomes in women. In this cross-sectional study, we determined the current prevalence of M. genitalium infection and the rate of macrolide resistance in M. genitalium isolates, in patients visiting two large Dutch sexually transmitted infection (STI) clinics, to evaluate whether the recommendations in Dutch guidelines should be revised. In addition, risk factors for M. genitalium were identified. In total, 3225 patients were included. M. genitalium prevalence rates were 13.8% for all patients; 20.1% for men who have sex with men, 8.2% for men who have sex with women, and 12.6% for women. Macrolide resistance-associated mutations were detected in 66% of the patients infected with M. genitalium. Age, educational level, country of origin, number of sexual partners, HIV-positivity, infection with Neisseria gonorrhoeae, and urethral symptoms in men were independently associated with M. genitalium infection. In conclusion, we found very high prevalence rates and macrolide resistance rates of M. genitalium in patients visiting STI clinics.

Validation of an algorithm for semiautomated surveillance to detect deep surgical site infections after primary total hip or knee arthroplasty-A multicenter study.

OBJECTIVE: Surveillance of healthcare-associated infections is often performed by manual chart review. Semiautomated surveillance may substantially reduce workload and subjective data interpretation. We assessed the validity of a previously published algorithm for semiautomated surveillance of deep surgical site infections (SSIs) after total hip arthroplasty (THA) or total knee arthroplasty (TKA) in Dutch hospitals. In addition, we explored the ability of a hospital to automatically select the patients under surveillance. DESIGN: Multicenter retrospective cohort study. METHODS: Hospitals identified patients who underwent THA or TKA either by procedure codes or by conventional surveillance. For these patients, routine care data regarding microbiology results, antibiotics, (re)admissions, and surgeries within 120 days following THA or TKA were extracted from electronic health records. Patient selection was compared with conventional surveillance and patients were retrospectively classified as low or high probability of having developed deep SSI by the algorithm. Sensitivity, positive predictive value (PPV), and workload reduction were calculated and compared to conventional surveillance. RESULTS: Of 9,554 extracted THA and TKA surgeries, 1,175 (12.3%) were revisions, and 8,378 primary surgeries remained for algorithm validation (95 deep SSIs, 1.1%). Sensitivity ranged from 93.6% to 100% and PPV ranged from 55.8% to 72.2%. Workload was reduced by ≥98%. Also, 2 SSIs (2.1%) missed by the algorithm were explained by flaws in data selection. CONCLUSIONS: This algorithm reliably detects patients with a high probability of having developed deep SSI after THA or TKA in Dutch hospitals. Our results provide essential information for successful implementation of semiautomated surveillance for deep SSIs after THA or TKA.

Evaluating the prevalence and risk factors for macrolide resistance in Mycoplasma genitalium using a newly developed qPCR assay.

Mycoplasma genitalium (MG) is a sexually transmitted bacterium in which macrolide resistance is rapidly increasing, limiting treatment options. We validated a new assay to detect the presence of macrolide resistance-associated mutations in MG (MG-MRAM). In 2018, symptomatic and asymptomatic clients visiting sexually transmitted infections (STI) clinics in Amsterdam or The Hague were tested for MG using transcription mediated amplification (TMA) assays. The sensitivity to detect MG of the newly developed MG-MRAM qPCR was compared to the MgPa qPCR, both in relation to the TMA assay. For the sensitivity and specificity to detect relevant mutations the MG-MRAM qPCR was compared to 23SrRNA sequencing analysis. The qPCR was subsequently used to determine the presence of MG-MRAM at different anatomical locations and to identify risk factors for MG-MRAM. MG-positive clients (402) providing 493 MG-positive samples were included. In total 309/493 (62.7%) samples from 291 (72.4%) clients were successfully typed with the MG-MRAM qPCR. The MG-MRAM qPCR had a sensitivity of 98.6% (95%CI 91.1%-99.9%) and specificity of 94.1% (95%CI 78.9%-99.0%) to detect MG-MRAM compared to sequencing analysis. Infection with MG-MRAM was detected in 193/291 (66.3%) clients: in 129/178 (72.5%) men and 64/113 (56.6%) women (p = 0.005). Prevalence of MG-MRAM was significantly higher in men, clients with a higher education, HIV-positive clients and clients with >10 sexual partners in the previous six months, but in multivariable analysis no factor was significantly associated with MG-MRAM presence. Since MG-MRAM prevalence was very high, testing for MG-MRAM is essential if treatment for MG is considered, and can be performed with this sensitive and specific qPCR test in routine diagnostics.

INNO-LiPA DNA line probe assay misidentification of M. smegmatis as Mycobacterium fortuitum complex.

Seven weeks after being kicked in the face by a cow, a 34-year-old male patient developed a posttraumatic mycobacterial lymphadenitis. A rapidly growing mycobacterial isolate cultured from a surgically drained lymphadenitis pus specimen was identified as Mycobacterium smegmatis by matrix-assisted laser desorption/ionization mass spectrometry and a combination of ITS-, hsp65-, and 16S rRNA-DNA sequence analysis, but as Mycobacterium fortuitum complex using the commercial INNO-LiPA Mycobacteria v2 line probe assay. As it is unclear if the misidentification of this strain is an exception, more research is required.

Preventing Staphylococcus aureus bacteremia and sepsis in patients with Staphylococcus aureus colonization of intravascular catheters: a retrospective multicenter study and meta-analysis.

Two previous studies in tertiary care hospitals identified Staphylococcus aureus colonization of intravascular (IV) catheters as a strong predictor of subsequent S. aureus bacteremia (SAB), even in the absence of clinical signs of systemic infection. Bacteremia was effectively prevented by timely antibiotic therapy. We conducted this study to corroborate the validity of these findings in non-university hospitals.Using the laboratory information management systems of the clinical microbiology departments in 6 Dutch hospitals, we identified patients who had IV catheters from which S. aureus was cultured between January 1, 2003, and December 31, 2008. Patients with demonstrated SAB between 7 days before catheter removal and 24 hours after catheter removal were excluded. We extracted clinical and demographic patient data from the patients' medical records. The primary risk factor was initiation of anti-staphylococcal antibiotic therapy within 24 hours, and the primary endpoint was SAB >24 hours after IV catheter removal. Subsequently, we performed a systematic review and meta-analysis of all observational studies evaluating the effect of antibiotic therapy for S. aureus IV catheter tip colonization.In the current study, 18 of the 192 included patients developed subsequent SAB, which was associated with not receiving antibiotic therapy within 24 hours (odds ratio [OR], 4.2; 95% confidence interval [CI], 1.1-15.6) and with documented exit-site infection (OR, 3.3; 95% CI, 1.2-9.3). When we combined these results with results of a previous study in a university hospital, a third risk factor was also associated with subsequent SAB, namely corticosteroid therapy (OR, 2.9; 95% CI, 1.3-6.3). We identified 3 other studies, in addition to the present study, in a systematic review. In the meta-analysis of these studies, antibiotic therapy yielded an absolute risk reduction of 13.6% for subsequent SAB. The number needed to treat to prevent 1 episode of SAB was 7.4.We conclude that early initiation of antibiotic therapy for IV catheters colonized with S. aureus prevents subsequent SAB.