Nosocomial Methicillin-resistant Staphylococcus Aureus Isolates Research Articles

806. Uncluttering Case Clusters: Use of Rapid Sequencing to Exclude Transmission Events

Abstract Background Newly identified multi-drug resistant organisms (MDRO) isolated from hospitalized patients with shared epidemiological characteristics can either represent transmission events or independent, unrelated acquisitions. Whole genome sequencing (WGS) can improve the efficiency of investigations triggered by MDRO cases with apparent epidemiological linkages by early exclusion of clonality. We report an implementation of WGS to investigate a cluster of methicillin-resistant Staphylococcus aureus (MRSA) and a cluster of carbapenem-resistant Enterobacterales (CRE) inpatient nosocomial infections. Methods Study participants included five Neonatal ICU (NICU) patients with nosocomial MRSA isolates recovered between June and August 2020, and two Respiratory Acute Care Unit (RACU) patients with nosocomial CRE infections in October 2020. Routine unit surveillance activities and characterization using standard epidemiologic criteria identified the isolates as nosocomial to their respective unit. The isolates then underwent WGS and single nucleotide polymorphism (SNP)-based relatedness analysis. Results The MRSA cluster included five neonates with either clinical or surveillance isolates. WGS identified one of the five isolates as methicillin susceptible S. aureus due to the absence of the mecA or mecC resistance gene, despite growth on chromogenic MRSA screening agar. WGS revealed each of the five isolates as belonging to a distinct multi locus sequence type (MLST) group with thousands of SNP differences between samples. The CRE cluster included two patients with Klebsiella pneumoniae isolated from clinical cultures within five days of each other with identical antimicrobial susceptibility profiles. WGS of the two isolates revealed that they belonged to different MLSTs and had tens of thousands of differing SNPs. WGS results suggest that a nosocomial transmission linking these infections was highly unlikely. For both investigations, WGS returned results within thirty-six hours of sample receipt. Figure 1. Corynebacterium striatum Respiratory Cultures January 2020-February 2021 Panel showing single nucleotide polymorphism (SNP) differences between isolates for respective clusters. Conclusion Apparent transmission events can be resource intensive to investigate and manage. The application of rapid WGS allowed for early discontinuation of cluster investigations and conservation of resources. Disclosures Virginia M. Pierce, MD, UpToDate, Inc. (Other Financial or Material Support, Author) Mohamad Sater, PhD, Day Zero Diagnostics (Employee, Shareholder) Miriam Huntley, PhD, Day Zero Diagnostics (Employee, Shareholder) Ian Herriott, BS, Day Zero Diagnostics (Employee, Shareholder) Tim Farrell, MS, Day Zero Diagnostics, Inc. (Employee, Shareholder) David C. Rosenberg, MD, Cepheid Diagnostics (Consultant)Day Zero Diagnostics (Consultant)SeLux Diagnostics (Consultant) Erica S. Shenoy, MD, PhD, Vertex Pharmaceuticals (Individual(s) Involved: Self): I gave a single lecture in 3/2020 for which I received financial compensation, Other Financial or Material Support

The discontinuation of contact precautions for methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus: Impact upon patient adverse events and hospital operations

BackgroundContact precautions for endemic methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE) are a resource-intensive intervention to reduce healthcare-associated infections, potentially impeding patient throughput and limiting bed availability to isolate...

Extensive Dissemination of Methicillin-Resistant Staphylococcus aureus (MRSA) between the Hospital and the Community in a Country with a High Prevalence of Nosocomial MRSA

According to the EARS-Net surveillance data, Portugal has the highest prevalence of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) in Europe, but the information on MRSA in the community is very scarce and the links between the hospital and community are not known. In this study we aimed to understand the events associated to the recent sharp increase in MRSA frequency in Portugal and to evaluate how this has shaped MRSA epidemiology in the community. With this purpose, 180 nosocomial MRSA isolates recovered from infection in two time periods and 14 MRSA isolates recovered from 89 samples of skin and soft tissue infections (SSTI) were analyzed by pulsed-field gel electrophoresis (PFGE), staphylococcal chromosome cassette mec (SCCmec) typing, spa typing and multilocus sequence typing (MLST). All isolates were also screened for the presence of Panton Valentine leukocidin (PVL) and arginine catabolic mobile element (ACME) by PCR. The results showed that ST22-IVh, accounting for 72% of the nosocomial isolates, was the major clone circulating in the hospital in 2010, having replaced two previous dominant clones in 1993, the Iberian (ST247-I) and Portuguese (ST239-III variant) clones. Moreover in 2010, three clones belonging to CC5 (ST105-II, ST125-IVc and ST5-IVc) accounted for 20% of the isolates and may represent the beginning of new waves of MRSA in this hospital. Interestingly, more than half of the MRSA isolates (8/14) causing SSTI in people attending healthcare centers in Portugal belonged to the most predominant clones found in the hospital, namely ST22-IVh (n = 4), ST5-IVc (n = 2) and ST105-II (n = 1). Other clones found included ST5-V (n = 6) and ST8-VI (n = 1). None of the MRSA isolates carried PVL and only five isolates (ST5-V-t179) carried ACME type II. The emergence and spread of EMRSA-15 may be associated to the observed increase in MRSA frequency in the hospital and the consequent spillover of MRSA into the community.

Rapid and high-resolution distinction of community-acquired and nosocomial Staphylococcus aureus isolates with identical pulsed-field gel electrophoresis patterns and spa types

Methicillin-resistant Staphylococcus aureus (MRSA) represent a serious threat for public health worldwide. Of particular concern is the emergence of community-acquired MRSA, which is often difficult to distinguish from nosocomial MRSA due to a lack of suitable typing methods for early detection. For example, the USA300 pulsed-field gel electrophoresis (PFGE) pattern includes both the ‘classical’ community-acquired USA300 clone with spa type t008 and an epidemiologically unrelated nosocomial clone with spa type t024. Likewise, spa typing cannot distinguish the classic USA300 from nosocomial MRSA with the spa type t008. Since the fast and high-resolution distinction of these S. aureus types is important for infection prevention and surveillance, we investigated whether multiple-locus variable number tandem repeat fingerprinting (MLVF) can be applied to overcome these limitations. Indeed, MLVF correctly grouped 91 MRSA isolates belonging to the classic USA300 lineage, nosocomial MRSA isolates with the USA300 PFGE profile and spa type t024, and nosocomial MRSA isolates with spa type t008 into 3 distinct clusters. Importantly, several sub-clusters were also identified, reflecting epidemiological relationships between the respective isolates. We conclude that MLVF has the discriminatory power needed to rapidly distinguish very similar community-acquired and nosocomial MRSA isolates and that MLVF-based sub-clustering of isolates is highly useful for epidemiological investigations, outbreak prevention, and control.

Subinhibitory fluoroquinolone exposure selects for reduced beta-lactam susceptibility in methicillin-resistant Staphylococcus aureus and alterations in the SOS-mediated response

Growth of fluoroquinolone (FQ)-susceptible methicillin-resistant Staphylococcus aureus (MRSA) in the presence of sub-MIC FQ has been shown to enhance methicillin resistance in traditional nosocomial MRSA isolates. We aimed to confirm this phenomenon in nine community-associated MRSA (CA-MRSA) clinical isolates, and to identify candidate genes that might account for this unusual phenotype. Overnight growth of CA-MRSA strains in tryptic soy broth containing a subinhibitory concentration of either ciprofloxacin or levofloxacin resulted in a concentration-related increase in the number of colonies that grew on nafcillin agar, such that about one CFU in four exhibited significantly higher resistance to nafcillin, with only a modest increase in FQ MIC. No mutations were found in the quinolone-resistance determining region of gyrA and grlA. DNA microarray studies of a representative levofloxacin-exposed clone found that gene expression was increased for 53 open reading frames (ORFs), including norR and mecA, and decreased for 10. The majority of these ORFs encode regulatory and stress response proteins. In conclusion, sublethal exposure to FQ alters the SOS response in CA-MRSA and selects in a non-lethal manner for stable mutants with enhanced expression of methicillin resistance.

The role of molecular methods in the prevention of nosocomial methicillin-resistant Staphylococcus aureus clusters in cancer patients

In 2002, an increased incidence of nosocomial methicillin-resistant Staphylococcus aureus (MRSA) in our institution triggered a conventional investigation that failed to identify a common source. Molecular typing of the 70 nosocomial MRSA isolates obtained identified a predominant health care-associated clone A in the first trimester. Aggressive infection control measures led to a significant decrease in the number of isolates per 10,000 hospital days between the first trimester and the last 2 trimesters of 2003 (6.4 vs 3.8; P = .04). This was attributed to a decrease in clone A: SCCmec II, USA100, PVL gene-negative (2.3 per 10.000 patient-days vs 0.1 per 10,000 patient-days; P = .004). However, in 2003, 23% of the nosocomial isolates were SCCmec IV, USA300, PVL gene-positive. At that time, molecular methods allowed the detection and prevention of a nosocomial MRSA outbreak caused by a health care-associated clone; however, the community strains (SCCmec IV) have become a frequent cause of nosocomial infection in our institution.

Longitudinal analysis of chlorhexidine susceptibilities of nosocomial methicillin-resistant Staphylococcus aureus isolates at a teaching hospital in Taiwan

Chlorhexidine has been widely used for hand hygiene to prevent transmission of nosocomial pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). However, data on longitudinal surveillance of the susceptibility of MRSA isolates to chlorhexidine are limited. A total of 240 nosocomial MRSA isolates obtained in 1990, 1995, 2000 and 2005 at National Taiwan University Hospital (NTUH), a hospital where chlorhexidine gluconate was used for hand hygiene for more than 20 years, were included in the study. Chlorhexidine susceptibility, molecular typing using multilocus sequence typing and distribution of the qacA/B gene of these MRSA isolates were studied. The proportion of tested MRSA with a high MIC of chlorhexidine (>or=4 mg/L) was 1.7% in 1990, 50% in 1995, 40% in 2000 and 46.7% in 2005. Among these 83 isolates with high chlorhexidine MICs, 55.4% carried the qacA/B gene. MRSA isolates carrying the qacA/B gene were first detected in 1995 and belonged to a single clone at that time. However, the qacA/B gene was detected in MRSA isolates belonging to seven different clones in 2005. The proportion of tested MRSA isolates with high chlorhexidine MICs at NTUH increased from 1990 to 1995 and remained steady thereafter. The presence of the qacA/B gene may contribute to the spread of specific MRSA clones.

Spread of a single multiresistant methicillin-resistant Staphylococcus aureus clone carrying a variant of staphylococcal cassette chromosome mec type III isolated in a university hospital

The purpose of the study was the molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) isolates cultured from patients treated in seven wards of a university hospital in Lublin, Poland, over a 14-month period. Eleven nosocomial MRSA isolates were analyzed. Phenotypic identification of the isolates as MRSA was confirmed by the detection of the nuc and mecA genes using a multiplex PCR assay. The MRSA isolates were further characterized by pulsed-field gel electrophoresis, 16S-23S rRNA spacer length polymorphism analysis, and the simplex and multiplex SCCmec PCR assays. The MRSA isolates were found to be multiresistant: in addition to resistance to beta-lactam agents, they demonstrated resistance to ciprofloxacin, tetracycline, erythromycin, and gentamicin. The MRSA isolates were genetically identical and shared common pulsed-field gel electrophoresis profiles and 16S-23S rRNA spacer length polymorphism profiles. The PCR-based method revealed that the profile of the Lublin clone was identical to that of the Brazilian pandemic MRSA isolates. By SCCmec typing, all MRSA isolates harbored the C variant of the SCCmec type III that differed from the typical SCCmec type III pattern by the lack of locus F (414 bp). The results of this study indicate the spread of a single, multiresistant, MRSA clone in various wards of a university hospital over a 14-month period. The SCCmec structure harbored by the Lublin clone has previously been identified among Polish MRSA isolates representing the HoMRSA-Pol1 clone. The data from this study indicate that the Lublin MRSA clone is most probably genetically related to the HoMRSA-Pol1 clone. Moreover, this latter clone belongs to ST239, the same sequence type as the Hungarian and Brazilian pandemic MRSA isolates.

Fifteen-Year Study of the Changing Epidemiology of Methicillin-Resistant Staphylococcus aureus

Purpose The study’s purpose was to elucidate the evolutionary, microbiologic, and clinical characteristics of methicillin-resistant Staphylococcus aureus (MRSA) infections. Methods MRSA cases from military medical facilities in San Diego, from 1990 to 2004, were evaluated and categorized as community-acquired or nosocomial. Sequence type, staphylococcal chromosomal cassette gene type, and Panton-Valentine leukocidin gene status were determined for a subset of isolates. Results Over the 15-year period, 1888 cases of MRSA were identified; 65% were community acquired. The incidence (155 infections/100 000 person-year in 2004) and household-associated cases rapidly increased since 2002. Among persons with community-acquired MRSA, 16% were hospitalized and only 17% were initially given an effective antibiotic. Community-acquired MRSA cases compared with nosocomial MRSA cases were more often soft-tissue and less often urinary, lung, or bloodstream infections ( P < .001). Patients with community-acquired MRSA were younger (22 vs 64 years, P < .001) and less likely to have concurrent medical conditions (9% vs 98%, P < .001). Clindamycin resistance increased among community-acquired MRSA isolates during 2003 and 2004 compared with previous years (79% vs 13%, P < .001). Genetically, nosocomial MRSA isolates were significantly different than those acquired in the community. Although community-acquired MRSA isolates were initially diverse by 2004, one strain (staphylococcal chromosomal cassette type IV, sequence type 8, Panton-Valentine leukocidin gene positive) became the predominant isolate. Conclusions Community-acquired and intrafamilial MRSA infections have increased rapidly since 2002. Our 15 years of surveillance revealed the emergence of distinct community-acquired MRSA strains that were genetically unrelated to nosocomial MRSA isolates from the same community.

Risk factors for specific methicillin-resistant Staphylococcus aureus clones in a Korean hospital

To analyse the risk factors for nosocomial methicillin-resistant Staphylococcus aureus (MRSA) infections caused by different clonal types. A total of 134 non-duplicate nosocomial MRSA isolates were analysed for clonal types by molecular typing techniques. The medical records of 90 patients who had documented MRSA infection were evaluated retrospectively. Two predominant MRSA clones of sequence types (STs) ST239 (n = 75) and ST5 (n = 39) accounted for 85% of the isolates. Management of patients in the departments of orthopaedic surgery, neurosurgery and plastic surgery was identified as a risk factor for infection with MRSA of ST239, while the presence of intravascular catheters was a risk factor for infection with ST5. Pulmonary infection was significantly higher in the patients infected with ST239 strains than in the patients infected with ST5 strains (P < 0.05). The overall mean duration of antimicrobial therapy for the patients with ST239 infection was significantly more than that for the patients with ST5 infection (P < 0.05). ST239 and ST5 were the predominant MRSA clones in the study hospital. Risk factors were significantly different between ST239 and ST5 strains. The results of this study will be of use in designing larger prospective epidemiological studies for MRSA infection based on clonal types.

Epidemiology of Methicillin-ResistantStaphylococcus aureusat a University Hospital in the Canary Islands

To describe the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) at a university hospital in Tenerife, Canary Islands, during a 40-month period and to evaluate the effectiveness of the application of control measures. Laboratory-based surveillance, medical charts and microbiological records review, and characterization of strains by pulsed-field gel electrophoresis (PFGE) were used to describe the epidemiology. Infection control practices were introduced as an intervention. A 650-bed, tertiary-care university hospital. Patients with clinical and nasal isolates of MRSA and colonized staff members. The rate of nosocomial MRSA infections was 32.5% for 1997, 17.9% for 1998, 14.5% for 1999, and 25.6% during the first 4 months of 2000. The major sites of isolation for nosocomial MRSA infection included surgical wounds (25%) and the lower respiratory tract (24%). Intensive care units and surgical specialties had more frequent MRSA cases. Characteristics associated with nosocomial MRSA isolates included prior use of intensive antibiotic therapy, prolonged hospital stays, major underlying illness, invasive procedures, and older age. PFGE type A (subtype A1) was the strain most frequently found and the only PFGE type involved in clusters. Surveillance cultures and contact droplet precautions were followed by decreased rates for 2 years. Nevertheless, the spread of PFGE subtype A1 to many different areas of the hospital and the increase in incidence during the first third of 2000 indicates either that surveillance cultures were not used widely enough or that compliance with isolation measures was suboptimal.

Community-acquired methicillin-resistant Staphylococcus aureus in a rural American Indian community.

Until recently, methicillin-resistant Staphylococcus aureus (MRSA) infections have been acquired primarily in nosocomial settings. Four recent deaths due to MRSA infection in previously healthy children in the Midwest suggest that serious MRSA infections can be acquired in the community in rural as well as urban locations. To document the occurrence of community-acquired MRSA infections and evaluate risk factors for community-acquired MRSA infection compared with methicillin-susceptible S aureus (MSSA) infection. Retrospective cohort study with medical record review. Indian Health Service facility in a rural midwestern American Indian community. Patients whose medical records indicated laboratory-confirmed S aureus infection diagnosed during 1997. Proportion of MRSA infections classified as community acquired based on standardized criteria; risk factors for community-acquired MRSA infection compared with those for community-acquired MSSA infection; and relatedness of MRSA strains, determined by pulsed-field gel electrophoresis (PFGE). Of 112 S aureus isolates, 62 (55%) were MRSA and 50 (45%) were MSSA. Forty-six (74%) of the 62 MRSA infections were classified as community acquired. Risk factors for community-acquired MRSA infections were not significantly different from those for community-acquired MSSA. Pulsed-field gel electrophoresis subtyping indicated that 34 (89%) of 38 community-acquired MRSA isolates were clonally related and distinct from nosocomial MRSA isolates found in the region. Community-acquired MRSA may have replaced community-acquired MSSA as the dominant strain in this community. Antimicrobial susceptibility patterns and PFGE subtyping support the finding that MRSA is circulating beyond nosocomial settings in this and possibly other rural US communities.

Methicillin-Resistant Staphylococcus aureus as a Community Organism

An increase in methicillin-resistant Staphylococcus aureus (MRSA) infections prompted a study of MRSA during a 21-month period in a 600-bed university hospital in southern Texas. MRSA cases were classified as community, nosocomial, or transfer cases. A case-control study of risk factors for community MRSA compared with community methicillin-susceptible S. aureus (MSSA) was performed. Pulsed field gel electrophoresis (PFGE) of whole cell DNA typing was used as a marker of strain identity for 31 consecutive isolates collected during the last 8 months of the study. During the 21 months there were 170 patients with MRSA infection or colonization, an incidence of 0.2 per 1,000 patient-days. Ninety-nine (58%) of 170 isolates were from community cases; the community to nosocomial case ratio was 2:1. No significant risk factors differentiated patients with community MRSA compared with community MSSA. Most community MRSA isolates studied (15 [68%] of 22) had distinct PFGE patterns, as did many nosocomial MRSA isolates (4 [44%] of 9). MRSA isolates were commonly present on admission to the hospital, and multiple MRSA strains were demonstrated among both community and hospital isolates.