Annet Troelstra

Evidence for in-vivo transfer of mecA DNA between strains of Staphylococcus aureus

Summary Staphylococcus aureus is thought to have acquired mecA DNA by horizontal transfer. DNA fingerprints made by restriction nucleases that cut certain sequences of DNA are used to compare complete genomes or particular genes between bacteria. We isolated an epidemic mecA − meticillin-susceptible S aureus genotype and, subsequently, a rare isogeneic mecA + meticillin-resistant S aureus (MRSA) genotype from a neonate who had never been in contact with MRSA. This MRSA contained mecA DNA that was identical to that in a coagulase-negative staphylococcal strain isolated from this patient, but different from other MRSA genotypes. We believe that this MRSA was formed in vivo by horizontal transfer of the mecA DNA between two staphylococcal species.

Role of healthcare workers in outbreaks of methicillin-resistant Staphylococcus aureus: a 10-year evaluation from a Dutch university hospital.

BACKGROUND AND OBJECTIVE The benefit of screening healthcare workers (HCWs) at risk for methicillin-resistant Staphylococcus aureus (MRSA) carriage and furloughing MRSA-positive HCWs to prevent spread to patients is controversial. We evaluated our MRSA program for HCWs between 1992 and 2002. SETTING A university medical center in The Netherlands, where methicillin resistance has been kept below 0.5% of all nosocomial S. aureus infections using active surveillance cultures and isolation of colonized patients. DESIGN HCWs caring for MRSA-positive patients or patients in foreign hospitals were screened for MRSA. MRSA-positive HCWs had additional cultures, temporary exclusion from patient-related work, assessment of risk factors for persisting carriage, decolonization therapy with mupirocin intranasally and chlorhexidine baths for skin and hair, and follow-up cultures. RESULTS Fifty-nine HCWs were colonized with MRSA. Seven of 840 screened employees contracted MRSA in foreign hospitals; 36 acquired MRSA after contact with MRSA-positive patients despite isolation precautions (attack rate per outbreak varied from less than 1% to 15%). Our hospital experienced 17 MRSA outbreaks, including 13 episodes in which HCWs were involved. HCWs were index cases of at least 4 outbreaks. In 8 outbreaks, HCWs acquired MRSA after caring for MRSA-positive patients despite isolation precautions. CONCLUSION Postexposure screening of HCWs allowed early detection of MRSA carriage and prevention of subsequent transmission to patients. Where the MRSA prevalence is higher, the role of HCWs may be greater. In such settings, an adapted version of our program could help prevent dissemination.

Transmissibility of livestock-associated methicillin-resistant Staphylococcus aureus (ST398) in Dutch hospitals

We quantified nosocomial transmission rates of sequence type (ST) 398 methicillin-resistant Staphylococcus aureus (MRSA) (an emerging livestock-associated MRSA clone) and non-ST398 MRSA isolates in patients hospitalized without infection control measures in 51 Dutch hospitals. Identification of 174 index patients initiated 139 post-exposure screenings of 9925 persons. There were 65 genotype-confirmed secondary cases (three and 62 for ST398 and non-ST398 MRSA, respectively), yielding a relative transmission risk for ST398 MRSA of 0.28 (95% CI 0.09-0.90), which was not sensitive to adjustment for duration of hospitalization at time of detection. Nosocomial transmission of ST398 MRSA is 72% less likely than that of non-ST398 MRSA strains.

Dual effects of soluble CD14 on LPS priming of neutrophils.

To evaluate the effect of soluble CD14 (sCD14) on human neutrophil response to lipopolysaccharide (LPS), we developed an LPS‐priming assay that measures the chemiluminescence response to N‐formyl‐methionyl‐leucyl‐phenylalanine stimulation. Priming by 1 ng/mL rough LPS occurred in the presence of either serum or recombinant LPS‐binding protein (LBP) only. Priming was completely CD14‐dependent because preincubation of the neutrophils with an anti‐CD14 monoclonal antibody prevented priming. We hypothesize that sCD14 enhances LPS response in neutrophils, but this response is not as effective as LPS response via membrane CD14 (mCD14). In our experiments sCD14 is present in an excess compared with mCD14. Priming of neutrophils occurs with low LBP, supposedly via sCD14‐LPS complexes. With high LBP, addition of sCD14 inhibited LPS‐priming of neutrophils. In that case, LPS may be transported to sCD14, preventing a more effective response via mCD14. In this study we demonstrate that the effect of sCD14 on neutrophil response to LPS is a delicate balance between activation and inhibition depending on concentration of serum or LBP. J. Leukoc. Biol. 61: 173–178; 1997.

Genotyping and Preemptive Isolation to Control an Outbreak of Vancomycin-Resistant Enterococcus faecium

BACKGROUND Control of vancomycin-resistant Enterococcus faecium (VRE) in European hospitals is hampered because of widespread asymptomatic carriage of VRE by healthy Europeans. In 2000, our hospital (The University Medical Center Utrecht, Utrecht, The Netherlands) was confronted with a large outbreak of VRE. INTERVENTION On the basis of genotyping (by pulsed-field gel electrophoresis), epidemic and nonepidemic VRE strains were distinguished, and infection-control measures were exclusively targeted toward epidemic VRE. The outbreak was retrospectively divided into 3 periods of different infection-control measures. Compliance with use of alcohol-based hand rubs was enforced during all periods. Period I involved active surveillance, isolation of carriers, and cohorting (duration, 4 months); preemptive isolation of high-risk patients for VRE colonization was added in period II (7 months); and cohorting and preemptive isolation were abandoned in period III (18 months). METHODS When the outbreak was identified, 27 patients in 6 wards were colonized; 93% were colonized with an epidemic VRE strain. Detection rates of nonepidemic VRE were 3.5%, 3.0%, and 2.9% among 683, 810, and 977 screened patients in periods I, II, and III, respectively, comparable to a prevalence of 2% (95% confidence interval [CI], 1%-3.5%) among 600 nonhospitalized persons. The relative risks of detecting epidemic VRE in periods II and III, compared with period I, were 0.67 (95% CI, 0.41-1.10) for period II and 0.02 (95% CI, 0.002-0.6) for period III. Infection-control measures were withheld for patients colonized with nonepidemic VRE (76 [54%] of 140 patients with a test result positive for VRE). Use of alcohol-based hand rubs increased by 31%-275% in outbreak wards. CONCLUSION Genotyping-targeted infection control, isolation of VRE carriers, enhancement of hand-hygiene compliance, and preemptive isolation successfully controlled nosocomial spread of epidemic VRE infection.

Methicillin-resistant Staphylococcus aureus carriage among patients after hospital discharge.

BACKGROUND AND OBJECTIVE At the University Medical Center Utrecht (UMCU), follow-up implies an inventory of risk factors and screening for MRSA colonization among all MRSA-positive patients for at least 6 months. If risk factors or positive cultures persist or re-emerge, longer follow-up is indicated and isolation at readmission. This study investigated how long MRSA-positive patients remained colonized after hospital discharge and which risk factors were important. Furthermore, the results of eradication therapy were evaluated. DESIGN All patients who were positive for MRSA at the UMCU between January 1991 and January 2001 were analyzed regarding carriage state, presence of risk factors for prolonged carriage of Staphylococcus aureus, and eradication treatment. RESULTS A total of 135 patients were included in the study. The median follow-up time was 1.2 years. Eighteen percent of the patients were dismissed from follow-up 1 year after discharge. Only 5 patients were dismissed after 6 months. Among patients with no risk factors, eradication treatment was effective for 95% within 1 year. Among patients with persistent risk factors, treatment was effective for 89% within 2 years. CONCLUSIONS Based on these findings, eradication therapy should be prescribed for all MRSA carriers, independent of the presence of risk factors. MRSA-positive patients should be evaluated for 6 months for the presence of risk factors and MRSA carriage. Screening for risk factors is important because intermittent MRSA carriage was found in a significant number of our patients. Patients with negative MRSA cultures and without risk factors for 12 months can be safely dismissed from follow-up.

Is Methicillin‐ResistantStaphylococcus aureusMore Contagious Than Methicillin‐SusceptibleS. aureusin a Surgical Intensive Care Unit? •

Background and Objective: In the Netherlands, the prevalence of methicillin resistance among Staphylococcus aureus isolates has been kept to less than 1% by using active screening programs and isolation. At the University Medical Center Utrecht (UMCU), an active screening program for methicillin-resistant S. aureus (MRSA) in the surgical intensive care unit (ICU) was implemented in 1986. Between 1992 and 2001, only 6 patients with MRSA were admitted to the surgical ICU. However, 4 of these 6 strains were able to spread to 23 other patients and 15 healthcare workers (HCWs). We were surprised by the epidemic behavior of these strains and wondered whether this was exceptional for S. aureus or whether methicillin-susceptible S. aureus (MSSA) was also spreading in the ICU. Design: A 2-month, prospective, observational study to investigate the incidence and spread of MSSA in the surgical ICU of UMCU and historical data collected during a 10-year period regarding MRSA. Setting: A 10-bed surgical ICU in a 1,042-bed teaching hospital. Results: Weekly swabs revealed the presence of MSSA in 11 (24%) of 45 patients and 16 (22%) of 72 HCWs. Of all 4,105 patient–HCW contacts, there were only 21 episodes in which both the patient and the HCW were found to carry MSSA. With the use of pulsed-field gel electrophoresis, no identical strains could be identified. Conclusion: In our surgical ICU, MRSA seems to spread more easily than MSSA, probably because of selection under antibiotic pressure or a still unknown intrinsic factor within MRSA.

Automated Surveillance for Healthcare-Associated Infections: Opportunities for Improvement

Surveillance of healthcare-associated infections is a cornerstone of infection prevention programs, and reporting of infection rates is increasingly required. Traditionally, surveillance is based on manual medical records review; however, this is very labor intensive and vulnerable to misclassification. Existing electronic surveillance systems based on classification algorithms using microbiology results, antibiotic use data, and/or discharge codes have increased the efficiency and completeness of surveillance by preselecting high-risk patients for manual review. However, shifting to electronic surveillance using multivariable prediction models based on available clinical patient data will allow for even more efficient detection of infection. With ongoing developments in healthcare information technology, implementation of the latter surveillance systems will become increasingly feasible. As with current predominantly manual methods, several challenges remain, such as completeness of postdischarge surveillance and adequate adjustment for underlying patient characteristics, especially for comparison of healthcare-associated infection rates across institutions.

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).

Prevalence of hospital-acquired infections during successive surveillance surveys conducted at a university hospital in the Netherlands.

OBJECTIVE To monitor hospital-wide trends in the prevalence of hospital-acquired infections (HAIs) in order to identify areas where the risk of infection is increasing. METHODS Successive surveillance surveys were conducted twice yearly, from November 2001 until May 2004, to determine the prevalence of HAIs at 2 Dutch hospitals, using Centers for Disease Control and Prevention criteria. RESULTS In all, 340 HAIs were observed in 295 (11.1%) of 2,661 patients surveyed. The overall prevalence per survey varied from 10.2% to 15.6%, with no significant differences between successive surveys. In the surgical department, the prevalence of HAIs increased from 10.8 cases per 100 surgeries in November 2001 to 20.4 cases per 100 surgeries in May 2002. Further analysis revealed a high prevalence of surgical site infection among patients who had an orthopedic procedure performed. In the neurology-neurosurgery department, the prevalence increased from 13.0 cases per 100 patients in May 2002 to 26.6 cases per 100 patients in May 2003 and involved several types of infection. Further analysis retrieved exceptionally high incidences of infections associated with cerebrospinal fluid drainage. Specific infection control interventions were developed and implemented in both departments. The total cost of the surveys was estimated to be euro9,100 per year. CONCLUSION Successive performance of surveillance surveys is a simple and cheap method to monitor the prevalence of infection throughout the hospital and appeared instrumental in identifying 2 departments with increased infection rates.