Successful containment of two vancomycin-resistant Enterococcus faecium (VRE) outbreaks in a Dutch teaching hospital using environmental sampling and whole-genome sequencing.

BACKGROUND: Vancomycin-resistant enterococci (VRE) may cause nosocomial outbreaks. This article describes all VRE carriers that were identified in 2018 at Elisabeth-Tweesteden Hospital, Tilburg, The Netherlands. AIM: To investigate the genetic relatedness of VRE isolates and the possibility of a common environmental reservoir using environmental sampling and whole-genome sequencing (WGS). METHODS: Infection control measures consisted of contact isolation, contact surveys, point prevalence screening, environmental sampling, cleaning and disinfection. VRE isolates were sequenced using a MiSeq sequencer (Illumina, San Diego, CA, USA), and assembled using SPAdes v.3.10.1. A minimal spanning tree and a neighbour joining tree based on allelic diversity of core-genome multi-locus sequence typing and accessory genes were created using Ridom SeqSphere+ software (Ridom GmbH, Münster, Germany). FINDINGS: Over a 1-year period, 19 VRE carriers were identified; of these, 17 were part of two outbreaks. Before environmental cleaning and disinfection, 55 (14%) environmental samples were VRE-positive. Fifty-one isolates (23 patient samples and 28 environmental samples) were available for WGS analysis. Forty-four isolates were assigned to ST117-vanB, five were assigned to ST17-vanB, and two were assigned to ST80-vanB. Isolates from Outbreak 1 (N=22) and Outbreak 2 (N=22) belonged to ST117-vanB; however, WGS showed a different cluster type with 257 allelic differences. CONCLUSION: WGS of two outbreak strains provided discriminatory information regarding genetic relatedness, and rejected the hypothesis of a common environmental reservoir. A high degree of environmental contamination was associated with higher VRE transmission. Quantification of environmental contamination may reflect the potential for VRE transmission and could therefore support the infection control measures.

Short-course aminoglycosides as adjunctive empirical therapy in patients with Gram-negative bloodstream infection, a cohort study.

OBJECTIVE: Short-course aminoglycosides as adjunctive empirical therapy to ß-lactams in patients with a clinical suspicion of sepsis are used to broaden antibiotic susceptibility coverage and to enhance bacterial killing. We quantified the impact of this approach on 30-day mortality in a subset of sepsis patients with a Gram-negative bloodstream infection. METHODS: From a prospective cohort study conducted in seven hospitals in the Netherlands between June 2013 and November 2015, we selected all patients with Gram-negative bloodstream infection (GN-BSI). Short-course aminoglycoside therapy was defined as tobramycin, gentamicin or amikacin initiated within a 48-hour time window around blood-culture obtainment, and prescribed for a maximum of 2 days. The outcome of interest was 30-day all-cause mortality. Confounders were selected a priori for adjustment using a propensity score analysis with inverse probability weighting. RESULTS: A total of 626 individuals with GN-BSI who received ß-lactams were included; 156 (24.9%) also received aminoglycosides for a median of 1 day. Patients receiving aminoglycosides more often had septic shock (31/156, 19.9% versus 34/470, 7.2%) and had an eight-fold lower risk of inappropriate treatment (3/156, 1.9% versus 69/470, 14.7%). Thirty-day mortality was 17.3% (27/156) and 13.6% (64/470) for patients receiving and not receiving aminoglycosides, respectively; yielding crude and adjusted odds ratios for 30-day mortality for patients treated with aminoglycosides of 1.33 (95% CI 0.80-2.15) and 1.57 (0.84-2.93), respectively. CONCLUSIONS: Short-course adjunctive aminoglycoside treatment as part of empirical therapy with ß-lactam antibiotics in patients with GN-BSI did not result in improved outcomes, despite better antibiotic coverage of pathogens.

Added diagnostic value of broad-range 16S PCR on periprosthetic tissue and clinical specimens from other normally sterile body sites.

AIMS: Evaluation of 16S PCR in addition to the standard culture to improve the pathogen detection rate in clinical specimens. METHODS AND RESULTS: Microbiological culture and direct 16S PCR was performed on specimens from suspected prosthetic joint infection patients (cohort-1) and on tissues and fluids from other normally sterile body sites (cohort-2). Based on clinical and microbiological data, the detection rate for both methods was assessed, assuming no superiority of either 16S PCR or culture. In cohort-1, 469 specimens were obtained. Culture was positive in 170 (36·2%) specimens, 16S PCR detected 70 (41·2%) of those pathogens. Additionally, 16S PCR detected pathogens in 13 of 299 (4·3%) culture-negative specimens. In cohort-2, pathogens were cultured in 52 of 430 (12·1%) specimens and 16S PCR revealed those pathogens in 32 (61·5%) specimens. 16S PCR detected pathogens in 31 of 378 (8·2%) culture-negative specimens. CONCLUSIONS: Overall, the yield with 16S PCR was low. For cohort-1 16S PCR detected pathogens in 4·3% of culture-negative specimens, where this was 8·2% for cohort-2. SIGNIFICANCE AND IMPACT OF THE STUDY: Although direct 16S PCR cannot replace culture, it may offer a valuable additional diagnostic option for detection of difficult to culture micro-organisms in culture-negative clinical specimens.

Extended-spectrum ß-lactamase (ESBL) polymerase chain reaction assay on rectal swabs and enrichment broth for detection of ESBL carriage.

BACKGROUND: Extended-spectrum ß-lactamase (ESBL) screening and contact precautions on patients at high risk for ESBL carriage are considered important infection control measures. Since contact precautions are costly and may negatively impact patient care, rapid exclusion of ESBL carriage and therefore earlier discontinuation of contact precautions are desired. AIM: In the present study, the performance of an ESBL polymerase chain reaction (PCR) targeting blaCTX-M genes was evaluated as a screening assay for ESBL carriage. METHODS: Two methods were assessed: PCR performed directly on rectal swabs and PCR on enrichment broth after incubation overnight. The reference standard was culture of ESBL-producing Enterobacteriaceae on selective agar after overnight enrichment and confirmation by the combination disc diffusion method. Microarray was used for discrepancy analysis. A secondary analysis was performed to evaluate the added value of including a blaSHV target in the PCR. FINDINGS: A total of 551 rectal swabs from 385 patients were included, of which 28 (5%) were ESBL positive in culture. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 86%, 98%, 67%, and 99%, respectively, for PCR directly on swabs, and 96%, 98%, 75%, and 100%, respectively, for PCR on enrichment broth. Adding a blaSHV target to the assay resulted in a lower PPV without increasing the sensitivity and NPV. CONCLUSION: Screening for ESBL by PCR directly on rectal swabs has a high negative predictive value, is up to 48h faster than traditional culture and therefore facilitates earlier discontinuation of contact precautions, thereby improving patient care and saving valuable resources in the hospital.

Diarrhoea in general practice: when should a Clostridium difficile infection be considered? Results of a nested case-control study.

Clostridium difficile infections (CDIs) are frequent in hospitals, but also seem to increase in the community. Here, we aim to determine the incidence of CDI in general practice and to evaluate current testing algorithms for CDI. Three Dutch laboratories tested all unformed faeces (12,714) for C. difficile when diagnostic testing (for any enteric pathogen) was requested by a general practitioner (GP). Additionally, a nested case-control study was initiated, including 152 CDI patients and 304 age and sex-matched controls. Patients were compared using weighted multivariable logistic regression. One hundred and ninety-four samples (1.5%) were positive for C. difficile (incidence 0.67/10,000 patient years). This incidence was comparable to that of Salmonella spp. Compared with diarrhoeal controls, CDI was associated with more severe complaints, underlying diseases, antibiotic use and prior hospitalization. In our study, GPs requested a test for C. difficile in 7% of the stool samples, thereby detecting 40% of all CDIs. Dutch national recommendations advise testing for C. difficile when prior antibiotic use or hospitalization is present (18% of samples). If these recommendations were followed, 61% of all CDIs would have been detected. In conclusion, C. difficile is relatively frequent in general practice. Currently, testing for C. difficile is rare and only 40% of CDI in general practice is detected. Following recommendations that are based on traditional risk factors for CDI, would improve detection of CDI.

Rapid clearance of Giardia lamblia DNA from the gut after successful treatment.

To assess the time it takes for a real-time PCR to become negative after treatment of a Giardia lamblia infection, we evaluated two consecutive follow-up samples from 75 infected patients. Approximately 1 week after treatment all samples tested negative, indicating rapid clearance of parasitic DNA after successful treatment.

Secular trends in nosocomial bloodstream infections: antibiotic-resistant bacteria increase the total burden of infection.

BACKGROUND: It is unknown whether rising incidence rates of nosocomial bloodstream infections (BSIs) caused by antibiotic-resistant bacteria (ARB) replace antibiotic-susceptible bacteria (ASB), leaving the total BSI rate unaffected. METHODS: We investigated temporal trends in annual incidence densities (events per 100 000 patient-days) of nosocomial BSIs caused by methicillin-resistant Staphylococcus aureus (MRSA), ARB other than MRSA, and ASB in 7 ARB-endemic and 7 ARB-nonendemic hospitals between 1998 and 2007. RESULTS: 33 130 nosocomial BSIs (14% caused by ARB) yielded 36 679 microorganisms. From 1998 to 2007, the MRSA incidence density increased from 0.2 to 0.7 (annual increase, 22%) in ARB-nonendemic hospitals, and from 3.1 to 11.7 (annual increase, 10%) in ARB-endemic hospitals (P = .2), increasing the incidence density difference between ARB-endemic and ARB-nonendemic hospitals from 2.9 to 11.0. The non-MRSA ARB incidence density increased from 2.8 to 4.1 (annual increase, 5%) in ARB-nonendemic hospitals, and from 1.5 to 17.4 (annual increase, 22%) in ARB-endemic hospitals (P < .001), changing the incidence density difference from -1.3 to 13.3. Trends in ASB incidence densities were similar in both groups (P = .7). With annual increases of 3.8% and 5.4% of all nosocomial BSIs in ARB-nonendemic and ARB-endemic hospitals, respectively (P < .001), the overall incidence density difference of 3.8 increased to 24.4. CONCLUSIONS: Increased nosocomial BSI rates due to ARB occur in addition to infections caused by ASB, increasing the total burden of disease. Hospitals with high ARB infection rates in 2005 had an excess burden of BSI of 20.6 per 100 000 patient-days in a 10-year period, mainly caused by infections with ARB.

Rapid diagnostic testing of methicillin-resistant Staphylococcus aureus carriage at different anatomical sites: costs and benefits of less extensive screening regimens.

Multiple body site screening and pre-emptive isolation of patients at risk for methicillin-resistant Staphylococcus aureus (MRSA) carriage are considered essential for control of nosocomial spread. The relative importance of extranasal screening when using rapid diagnostic testing (RDT) is unknown. Using data from a multicentre study evaluating BD GeneOhm™ MRSA PCR (IDI), Xpert MRSA (GeneXpert) and chromogenic agar, added to conventional cultures, we determined cost-effectiveness assuming isolation measures would have been based on RDT results of different hypothetical screening regimes. Costs per isolation day avoided were calculated for regimes with single or less extensive multiple site RDT, regimes without conventional back-up cultures and when PCR would have been performed with pooling of swabs. Among 1764 patients at risk, MRSA prevalence was 3.3% (n = 59). In all scenarios the negative predictive value is above 98.4%. With back-up cultures of all sites as a reference, the costs per isolation day avoided were €15.19, €30.83 and €45.37 with 'nares only' screening using chromogenic agar, IDI and GeneXpert, respectively, as compared with €19.95, €95.77 and €125.43 per isolation day avoided when all body sites had been screened. Without back-up cultures costs per isolation day avoided using chromogenic agar would range from €9.24 to €76.18 when costs per false-negative RDT range from €5000 up to €50 000; costs for molecular screening methods would be higher in all scenarios evaluated. In conclusion, in a low endemic setting chromogenic agar screening added to multiple site conventional cultures is the most cost-effective MRSA screening strategy.

Rapid screening of methicillin-resistant Staphylococcus aureus using PCR and chromogenic agar: a prospective study to evaluate costs and effects.

Pre-emptive isolation of suspected methicillin-resistant Staphylococcus aureus (MRSA) carriers is considered essential for controlling the spread of MRSA, but noncolonized patients will be isolated unnecessarily as a result of a delay in diagnosis of 3-5 days with conventional cultures. We determined costs per isolation day avoided, and incremental costs of rapid MRSA screening tests when added to conventional screening, but with decisions on isolation measures based on PCR results. A prospective multicentre study evaluating BD GeneOhm MRSA PCR (`IDI') (BD Diagnostics, San Diego, CA, USA), Xpert MRSA (`GeneXpert') (Cepheid, Sunnyvale, CA, USA) and chromogenic agar (MRSA-ID) (bioMérieux, Marcy-l'Etoile, France) was performed in 14 Dutch hospitals. Among 1764 patients at risk, MRSA prevalence was 3.3% (n=59). Duration of isolation was 19.7 and 16.1 h with IDI and GeneXpert, respectively, and would have been 30.0 and 76.2 h when based on chromogenic agar and conventional cultures, respectively. Negative predictive values (at a patient level) were 99.5%, 99.1% and 99.5% for IDI, GeneXpert and chromogenic agar, respectively. Numbers of isolation days were reduced by 60% and 47% with PCR-based and chromogenic agar-based screening, respectively. The cost per test was €56.22 for IDI, €69.62 for GeneXpert and €2.08 for chromogenic agar, and additional costs per extra isolation day were €26.34. Costs per isolation day avoided were €95.77 (IDI) and €125.43 (GeneXpert). PCR-based decision-making added €153.64 (IDI) and €193.84 (GeneXpert) per patient to overall costs and chromogenic testing would have saved €30.79 per patient. Rapid diagnostic testing safely reduces the number of unnecessary isolation days, but only chromogenic screening, and not PCR-based screening, can be considered as cost saving.

Evaluation of a chromogenic agar (MRSASelect) for the detection of meticillin-resistant Staphylococcus aureus with clinical samples in The Netherlands.

A novel chromogenic medium for the detection of meticillin-resistant Staphylococcus aureus (MRSA), MRSASelect (Bio-Rad), was evaluated with clinical samples in a public health laboratory in The Netherlands. In total, 3000 samples were tested in the period January to March 2005, including 972 nose, 972 throat, 968 perineum, and 88 wound or urine samples. Presumptive MRSA colonies appeared pink/mauve on the MRSASelect medium. The performance of MRSASelect medium was compared with the routine screening method. Evaluation of the colony morphology showed that all confirmed MRSA isolates grew as pink/mauve colonies. None of the white colonies were MRSA strains. The number of false-positive pink/mauve colonies increased after prolonged incubation from 20 to 48 h. The specificity decreased from 92 % after 20 h incubation to 89 % after 48 h incubation. In total 70 MRSA strains were isolated, 55 of which were detected by the MRSASelect medium and 55 were detected by the routine screening method. Sensitivity was 78.6 % for both test procedures, and specificities were 99.5 and 100 %, respectively for the MRSASelect medium and the routine screening method. The addition of an enrichment broth to the MRSASelect medium increased the number of MRSA strains detected by 12 %. In total, 18 patients were MRSA positive, 4 of these were detected by the MRSASelect medium only and 1 was detected by the routine screening method only. Sensitivity on patient level was 94.4 and 77.8 % for the MRSASelect medium and the routine screening method, respectively, while specificities were 99.7 and 99.0 %.

Identification and susceptibility testing of Staphylococcus aureus by direct inoculation from positive BACTEC blood culture bottles.

This study explored the possibility of combining direct inoculation of tube coagulase and DNase tests, and the VITEK2 system, from BACTEC blood culture bottles in order to achieve rapid identification and susceptibility testing of Staphylococcus aureus. All isolates were identified correctly as S. aureus or coagulase-negative staphylococci (CNS). Antimicrobial susceptibility testing with the VITEK2 system gave 99.6% correct category agreement, with 0.1% very major errors and 0.3% minor errors among S. aureus isolates, and 97.4% correct category agreement, with 0.9% very major errors and 1.7% minor errors among CNS isolates. The results suggested that direct identification and susceptibility testing is sufficiently accurate for immediate reporting.

[Optimalization of antibiotic policy in the Netherlands. VII. SWAB-guidelines for antimicrobial therapy in adults patients with infectious endocarditis]. / Optimaliseren van het antibioticabeleid in Nederland. VII. SWAB-richtlijnen voor antimicrobiële therapie bij volwassen patiënten met infectieuze endocarditis.

The Working Party on Antibiotic Policy (Dutch acronym: SWAB) has developed guidelines for in-hospital antimicrobial therapy of adult patients with infective endocarditis. The choice and the duration of antimicrobial therapy are determined by the infecting micro-organism, the sensitivity of this micro-organism to antimicrobial therapy, the location of the endocarditis (left- or right-sided) and the presence of intracardial prosthetic material. While waiting for the culture results, the antibiotic treatment of an infected native valve is benzylpenicillin (in cases which begin subacutely or have a long history) or flucloxacillin (cases which begin acutely, are fulminant or in i.v. drug users), and gentamicin. If a prosthetic valve is infected then treatment of choice is vancomycin and gentamicin. Further antibiotic treatment depends on the causative micro-organism. Streptococci, enterococci and staphylococci are the most frequently occurring of these.

[Diagnostic image (157). A man with fever and a 'tache noire' after a holiday in South Africa. Rickettsiosis, probably African tick-bite fever]. / Diagnose in beeld (157). Een man met koorts en een 'tache noire' na een vakantie in Zuid-Afrika. Rickettsiose, waarschijnlijk Afrikaanse tekenkoorts.

A 54-year-old man presented with fever and an eschar, probably caused by African tick-bite fever, contracted during a holiday in South Africa. He recovered rapidly after treatment with doxycyclin.

Optimisation of the antibiotic guidelines in The Netherlands. VII. SWAB guidelines for antimicrobial therapy in adult patients with infectious endocarditis.

The Working Party on Antibiotic Policy (Dutch acronym is SWAB) is a Dutch organisation that develops guidelines for in-hospital antimicrobial therapy of bacterial infectious diseases. This present guideline describes the antimicrobial treatment for adult patients with infective endocarditis. The choice and duration of antimicrobial therapy is determined by the infecting micro-organism, sensitivity of this micro-organism for antimicrobial therapy, location of the endocarditis, left-sided or right-sided, and presence of intracardial prosthetic material. In this guideline, the empirical therapy for endocarditis is discussed as well as the therapy for the most frequent causative organisms: streptococci, enterococci, staphylococci and HACEK micro-organisms.

Decolonization of methicillin-resistant Staphylococcus aureus using oral vancomycin and topical mupirocin.

The objective of this study was to assess the efficacy and safety of a short course of oral vancomycin and intranasal mupirocin ointment in the eradication of methicillin-resistant Staphylococcus aureus (MRSA) colonization. During an outbreak of MRSA, the colonized subjects received oral vancomycin and topical mupirocin. They were screened for MRSA 1, 3, 6 and 12 months after decolonization. A questionnaire was developed to evaluate the side-effects of oral vancomycin. Thirty-five subjects were treated. Clearance was achieved in all cases, in 24 (69%) subjects after one course of therapy. Twenty-eight (80%) subjects experienced some side-effects, including six (17%) who did not tolerate oral vancomycin. Although oral vancomycin, in combination with topical mupirocin, is effective in the elimination of MRSA colonization, there is a need for further studies to confirm our results and to evaluate the safety of oral vancomycin.

Infectious complications and antibiotic use in renal transplant recipients during a 1-year follow-up.

OBJECTIVE: To evaluate infectious complications and antibiotic use in 192 renal transplant recipients. METHODS: Infectious complications and antibiotic use were monitored in all patients receiving renal transplantation at our center from 1992 to 1997. Risk factors for infectious complications were evaluated. Transplants and patient survival were monitored. The follow-up period was 1 year. RESULTS: One-hundred and ninety-two patients received renal transplants during the study period. The mean duration of urethral catheterisation after transplantation was 10.5 days (SD = 5). Seventy-one per cent (n = 137) of patients had at least one infectious episode. In all, 284 infectious episodes were monitored. The most frequent infections were: urinary tract infections 61%, respiratory tract infections 8%, intra-abdominal infections 7%, and cytomegalovirus infection 8%. Escherichia coli and Enterococcus faecalis were the most frequently isolated microorganisms. Seventy-four per cent (n = 142) of patients received 314 antimicrobial courses (284 for therapy, and 30 for prophylaxis). Female gender and duration of urethral catheterisation were risk factors for urinary tract infection. Cytomegalovirus reactivation was associated with acute graft rejection and additional immunosuppressive therapy. Overall mortality was 4%. Infection-related mortality was 2.6%. Mortality was associated with Enterobacteriaceae in three patients, with Pseudomonas aeroginosa in one patient and with Enterococcus faecalis in one patient. CONCLUSIONS: The incidence of infectious complications remains high in renal transplant recipients. Most cases of mortality were associated with infections. Early removal of the urethral catheter to reduce the risk of urinary tract infections is recommended.

The risk of Strongyloides stercoralis transmission from patients with disseminated strongyloidiasis to the medical staff.

To assess the risk of Strongyloides stercoralis transmission from two patients with disseminated strongyloidiasis to medical staff who had been in close contact with the patients, blood and stool specimens were obtained from medical staff two to three months after close contact with the patients. Antibodies to S. stercoralis were determined in blood. Stool specimens were tested for parasites with three different procedures.Forty-one medical staff were included. Culture and stool examination were negative in all subjects. Serology was negative in all subjects but one who had a borderline titer without signs or symptoms of strongyloidiasis. No evidence of transmission of S. stercoralis from patients with disseminated strongyloidiasis to medical staff was found.