Semi-selective broth improves screening for methicillin-resistant Staphylococcus aureus.

OBJECTIVES: To evaluate two enrichment broths for methicillin-resistant Staphylococcus aureus (MRSA) detection and compare results with direct plating. METHODS: Swabs from 1224 patients were re-analysed for MRSA in a central laboratory (Münster) using six methods. Swabs were suspended in 0.5 mL of non-selective enrichment broth (NB) and vortexed. Aliquots of 100 microL were inoculated on/into: (I) ChromID MRSA agar; (II) Columbia sheep blood (5%) agar (BA) and ChromID MRSA; (III, IV) NB incubated overnight followed by plating on BA and ChromID MRSA; and (V, VI) a semi-selective broth containing cefoxitin and aztreonam (TSB-SSI) incubated overnight followed by plating on BA and ChromID MRSA. In III-VI, 100 microL of the enriched broth was plated on each agar. RESULTS: The combined MRSA-positive rate was 21.5%. MRSA isolates detected by each method were: TSB-SSI, n = 223; NB, n = 205; BA and ChromID MRSA, n = 203; ChromID MRSA alone, n = 183. TSB-SSI detected more positive throat samples than the comparators and significantly reduced methicillin-susceptible S. aureus (MSSA) growth. The maximum sensitivity obtained was only 85%, possibly due to the study design using pre-used swabs and dilution of swab material. For 997 samples, results from Münster were compared with initial results. Peripheral laboratories identified 172 MRSA compared with Münster where 186, 186 and 204 MRSA were found for direct plating, NB and TSB-SSI broth, respectively. CONCLUSIONS: TSB-SSI was superior to both NB and direct plating on ChromID MRSA and BA. Despite re-using swabs for the study, we showed that routine diagnostic screening could be significantly improved, using a semi-selective enrichment broth.

The search and destroy strategy prevents spread and long-term carriage of methicillin-resistant Staphylococcus aureus: results from the follow-up screening of a large ST22 (E-MRSA 15) outbreak in Denmark.

In the aftermath of a methicillin-resistant Staphylococcus aureus (MRSA) ST22 hospital outbreak, we investigated the prevalence of long-term carriage, the efficacy of MRSA decolonization treatment (DT) and the spread of MRSA to households of patients and healthcare workers (HCWs). Furthermore, we evaluated the efficacy of repeated DT in long-term MRSA carriers. Of 250 index persons (58 HCWs and 192 patients), 102 persons (19 HCWs and 83 patients) and 67 household members agreed to participate. Samples from all 169 persons were taken from the nose, throat, wounds and devices/catheters, and urine samples were additionally taken from index persons. Samples from companion animals (n = 35) were taken from the nostrils and anus. Environmental sites (n = 490) screened were telephone, television remote control, toilet flush handle, favourite chair and skirting board beside the bed. Sixteen (19%) patients and two household members, but no HCWs, were ST22-positive. The throat was the most frequent site of colonization. In a multivariate analysis, chronic disease (p <0.001) and pharyngeal carriage (p <0.001) were associated with long-term MRSA carriage. MRSA was found in the environments of four long-term carriers. All animals tested were negative. MRSA-positive households were decolonized using nasal mupirocin TID and daily chlorhexidine body and hair wash for 5 days. Pharyngeal MRSA carriers also received fucidic acid (500 mg TID) combined with rifampicin (600 mg BID) or clindamycin (600 mg BID) for 7 days. The home environment was cleaned on days 2 and 5. At the end of follow-up, ten of 16 long-term carriers and the two household contacts were MRSA-negative. In conclusion, decolonization of MRSA carriers is possible, but should include treatment of household members and the environment.

Emergence and characterization of community-associated methicillin-resistant Staphyloccocus aureus infections in Denmark, 1999 to 2006.

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) infections has changed worldwide. From being strictly nosocomial, MRSA is now frequently found as a community-associated (CA) pathogen. Denmark has been a low-prevalence country for MRSA since the mid-1970s but has in recent years experienced an increasing number of CA-MRSA cases. The aim of this study was to describe the emergence of CA-MRSA infections in Denmark. All Danish MRSA specimens and corresponding clinical data from 1999 to 2006 were investigated. Isolates were analyzed by antibiotic resistance and molecular typing and were assigned to clonal complexes (CC). Clinical data were extracted from discharge summaries and general practitioners' notes, from which assessments of community association were made for all infected cases. CA-MRSA cases constituted 29.4% of all MRSA infections (n = 1,790) and an increasing proportion of the annual numbers of MRSA infections during the study period. CA-MRSA was associated with a young age, skin and soft tissue infections, and non-Danish origin. Transmission between household members was frequently reported. Molecular typing showed >60 circulating clones, where 89.4% of the isolates belonged to five CC (CC80, CC8, CC30, CC5, and CC22), 81.2% carried staphylococcal cassette chromosome mec IV, and 163/244 (69.4%) were positive for Panton-Valentine leukocidin. Clinical and microbiological characteristics indicated that import of MRSA occurs frequently. Resistance to > or =3 antibiotic classes was observed for 48.8% of the isolates. The emergence of CA-MRSA in Denmark was caused by diverse strains, both well-known and new CA-MRSA strains. The results suggest multiple introductions of MRSA as an important source for CA-MRSA infections in Denmark.

Methicillin-resistant Staphylococcus aureus: risk factors associated with community-onset infections in Denmark.

The proportion of methicillin-resistant Staphylococcus aureus (MRSA) in Denmark has been below 1% for more than 30 years. However, a marked increase in community-onset MRSA (CO-MRSA) started in 2002. To identify possible risk factors for CO-MRSA infections, a nationwide case-control study was conducted in 2004. Cases (34) were patients with CO-MRSA infections; controls (87) were patients with community-onset methicillin-sensitive S. aureus infections (CO-MSSA). Demographic and clinical data and exposures to possible risk factors during the last 24 months were collected with a structured telephone-administered questionnaire. Skin and soft tissue were the predominant sites of infection, both for cases (68%) and for controls (60%). A large proportion of cases (26%) and controls (38%) had an underlying skin disease. The majority of cases (76%) and controls (61%) had received antibiotics within the last 6 months, and 51% and 31%, respectively, had been hospitalized within the previous year. In a multivariate analysis, non-Danish origin, defined as being from or having parents from outside Denmark, was the only independent risk factor for CO-MRSA infection (OR 30.5, 95% CI 3.6-257.3). Prior hospitalization for >7 days within the previous 6 months tended to be associated with CO-MRSA infection (OR 5.7, 95% CI 0.9-36.4). The predominant MRSA clones found in this study were CC80 (26%), CC8 (24%) and CC5 (18%). Resistance to three or more antimicrobial drug classes was seen in 47% of CO-MRSA isolates. Panton-Valentine leukocidin was found in 47% of CO-MRSA isolates. Apart from a non-Danish origin, CO-MRSA shared the same risk factors as CO-MSSA, which makes control a challenge.

Evaluation of four selective agars and two enrichment broths in screening for methicillin-resistant Staphylococcus aureus.

To evaluate methicillin-resistant Staphylococcus aureus detection, we tested in vitro four selective agars and two enrichment broths apart and in combination. Tryptone soya broth with salt, aztreonam, and cefoxitin appeared to be the most sensitive medium. This broth was superior to a phenol red mannitol broth with aztreonam and ceftizoxime.

Epidemiology of European community-associated methicillin-resistant Staphylococcus aureus clonal complex 80 type IV strains isolated in Denmark from 1993 to 2004.

In Europe, community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) infections have been caused predominantly by isolates belonging to the "European CA-MRSA" clone (sequence type 80, staphylococcal cassette chromosome mec type IV). In this study, the epidemiology of European CA-MRSA was investigated on a nationwide scale, covering the period from 1993 to 2004. Denmark has been a low-prevalence country regarding MRSA since the mid-1970s but has experienced an increase in the number of new MRSA cases in recent years. Our results show that European CA-MRSA contributed to this increase. The isolates primarily caused skin and soft tissue infections (SSTIs) in patients outside hospitals, and transmission between household members was the predominant mode of spread. Although some of the isolates were found in hospitalized patients, nosocomial transmission seemed likely in only one instance, pointing to endogenous infections as an important factor. Compared to the CA-MRSA clone most common in the United States (USA300), the European CA-MRSA clone seems less well adapted to persist in hospital environments. Patients with a recent history of travel or family relation to the Mediterranean or Middle East were highly overrepresented. The epidemiological data indicated that the European CA-MRSA isolates were introduced into Denmark on multiple occasions, paralleled by an increasing level of genetic diversity of the isolates found during the study period. European CA-MRSA has previously been described as a rather uniform clone. However, we found pronounced, diverse pulsed-field gel electrophoresis subtypes, staphylococcal protein A gene (spa) types, and susceptibility patterns.