Clinical isolates of Staphylococcus intermedius masquerading as methicillin-resistant Staphylococcus aureus

Staphylococcus intermedius is a zoonotic organism that can be associated with human disease. We report two separate cases of S. intermedius infection in which a false-positive rapid penicillin binding protein 2a latex test in conjunction with the phenotypic properties of beta-hemolysis and coagulase positivity allowed the clinical isolates to masquerade as methicillin-resistant Staphylococcus aureus. 16S rRNA gene sequencing and the absence of mecA revealed the strains to be methicillin-susceptible S. intermedius.     

Evaluation of different chromogenic media for the detection of methicillin-resistant Staphylococcus aureus CC398 in broilers

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has emerged in a wide variety of animal species, including poultry. The objective of this study was to evaluate three different chromogenic media for MRSA clonal complex (CC) 398 detection in broilers. On three Belgian poultry farms, 50 broiler chickens were sampled per farm from both nose shell and cloaca. All swab specimens were enriched and inoculated the following day on three chromogenic media: chromID MRSA (bioMérieux), Brilliance MRSA 2 Agar (Oxoid) and MRSASelect (Bio-Rad). ChromID had the highest isolation rates, yet, Brilliance MRSA 2 Agar demonstrated the highest relative sensitivity, while MRSASelect and Brilliance MRSA 2 Agar showed the highest relative specificity. A subset of MRSA isolates was confirmed to be CC398 by the polymerase chain reaction (PCR) targeting sau1-hsdS1. In conclusion, Brilliance MRSA 2 Agar outperformed MRSASelect and chromID MRSA for the detection of MRSA in broilers.     

New chromogenic identification and detection of Staphylococcus aureus and methicillin-resistant S. aureus

This paper describes a new chromogenic plate medium, CHROMagar Staph aureus (CHROMagar, Paris, France), for the identification of Staphylococcus aureus on the basis of colony pigmentation. The abilities of CHROMagar Staph aureus, thermostable nuclease (DNase), and mannitol salt agar (MSA) to identify S. aureus isolates (n = 114) and discriminate between S. aureus and coagulase-negative staphylococci (CoNS; n = 22) were compared. CHROMagar Staph aureus proved to be more sensitive and specific than DNase and MSA, allowing a reliable, simple, and rapid method for the identification of S. aureus isolates. All CoNS encountered in this study with the exception of S. chromogenes could be easily differentiated from S. aureus on this medium. The supplementation with 4 microgram of oxacillin or methicillin per ml allowed simple identification of methicillin resistance in hospital-acquired S. aureus strains which show multiple-drug resistance profiles. Community-acquired methicillin-resistant S. aureus strains showing non-multi-drug resistance profiles require further evaluation on this new chromogenic medium. Methicillin or oxacillin resistance of all S. aureus isolates was confirmed by the detection of penicillin-binding protein 2a, encoded by the mecA gene, using the latex slide agglutination MRSA-Screen test (PBP 2' Test, DR900M; Oxoid).     

Methicillin-resistant Staphylococcus aureus detection using chromogenic media: the Sheffield experience

Methicillin-resistant Staphylococcus aureus (MRSA) continues to cause major problems, both in hospitals and the community. Microbiology departments need to review their methodology regularly to ensure that they are contributing in the most appropriate manner to the battle against MRSA. Media employing chromogenic enzymes to aid the isolation and identification of MRSA is a relatively new approach. In this study, 192 swabs from 112 different patients were inoculated on two chromogen-containing media and four other commonly used solid MRSA media to determine which gave the appropriate combination of sensititivity, specificity and speed of result. Methicillin-resistant S. aureus was isolated on at least one of the six media from 102 of the 192 swabs. Both chromogenic media proved to be statistically significantly more sensitive than the other media after overnight incubation and had a sensitivity of 96% after 48 hours' incubation. The recent introduction of chromogen-containing MRSA media offers microbiology laboratories the opportunity to isolate and confirm the majority of MRSA infections/colonisations in 24 hours, which should result in better patient care. The possible slight increase in costs should not provide a valid excuse for using inferior methodologies.     

Development and evaluation of a chromogenic agar medium for methicillin-resistant Staphylococcus aureus

We describe here the development and evaluation of MRSA ID, a new chromogenic agar medium for the specific isolation and identification of methicillin-resistant Staphylococcus aureus (MRSA). We used S. aureus ID (bioMérieux, La Balme Les Grottes, France) and supplemented it with various antimicrobials, including cefoxitin, ciprofloxacin, oxacillin, and methicillin. Cefoxitin proved to be superior to the other antimicrobials for the selection of MRSA from other strains of S. aureus. MRSA ID (consisting of S. aureus ID supplemented with 4 mg of cefoxitin/liter) was evaluated by the use of 747 swabs from various clinical sites. All specimens were also cultured on CHROMagar MRSA and oxacillin resistance screening agar base (ORSAB) and in selective mannitol broth (SMB). A total of 85 MRSA strains were isolated by a combination of all methods. After 22 to 24 h of incubation, 80% of the MRSA strains were isolated as green colonies on MRSA ID, compared with 59 and 62% of the strains that were isolated as colored colonies on CHROMagar MRSA and ORSAB, respectively. After 48 h of incubation, 89, 72, and 78% of the MRSA strains were isolated on MRSA ID, CHROMagar MRSA, and ORSAB, respectively. Sixty-five percent of the strains were isolated by growth in SMB. The specificities of MRSA ID, CHROMagar MRSA, ORSAB, and SMB were 99.5, 99.3, 97.9, and 92.8%, respectively, after 22 to 24 h of incubation. We conclude that MRSA ID is a sensitive and specific medium for the isolation and identification of MRSA.     

Detection of nasal colonization methicillin-resistant Staphylococcus aureus: a prospective study comparing real-time genic amplification assay vs selective chromogenic media

In contrast to "classical" genic amplification, real-time genic amplification can be performed in every laboratory without the need of sophisticated isolation procedures. Moreover, real-time genic amplification allows an early detection of meticillin resistant Staphylococcus aureus colonization, 2 hours compared to 1 or 2 days for culture. OBJECTIVE: In order to assess the feasibility on Smartcycler of the IDI-MRSA real-time genic amplification assay in comparison with chromogenic media. METHODS: A prospective study has been initiated in July 2004: nasal swabs were taken from patients entering the ICU, vascular surgery, diabetology and geriatry wards. During a 4 months period, 682 specimens have been obtained from 508 patients. RESULTS: Sixty-four (9.3%) patients were positive by genic amplification and selective agar culture (CHROMagar MRSA, MRSASelect and/or ORSAB), 19 (2.9%) were positive by genic amplification only (3 of these patients were under antibiotic treatment); 572 specimens remained negative by both methods. The sensitivity and specificity of this assay were 100% and 96% respectively with a positive predictive value of 70% and negative predictive value of 100%. Initially 82 nasal specimens were unresolved (12%). 38 were resolved following a freeze-thaw cycle. Thus, 44 (6.4%) were unresolved specimens. Comparison between CHROMagar MRSA and MRSASelect showed a good correlation for the detection at 24 hours (5.5% and 5.6% respectively). These two chromogenic media allowed a much better detection of MRSA than ORSAB medium within 24H. CONCLUSION: The results obtained by the early real-time genic amplification for the detection of meticillin resistant Staphylococcus aureus are promising. Despite 6.4% amplification failure, we consider that IDI-MRSA real-time genic amplification assay represents a significant breakthrough in the detection of colonization.     

Comparison of three selective chromogenic media for Methicillin-Resistant Staphylococcus aureus detection

PURPOSE: Rapid detection of Methicillin-Resistant Staphylococcus aureus (MRSA) is of major importance in hospital hygiene. In order to reduce the response time from screening laboratories, new selective chromogenic media have been developed and marketed by major microbiology companies. In this context, an evaluation of their performances was needed. MATERIALS AND METHODS: Media produced by Bio-Rad Laboratories (MRSASelect), Becton Dickinson (CHROMagar MRSA) and bioMérieux (chromID MRSA) were studied by 203screening samples, 110 of which were MRSA positive. Each Stahylococcus aureus was identified by catalase detection, Staphytect Plus Dryspot latex agglutination test and free coagulase detection, in addition to mannitol fermentation and Voges-Proskauer tests in the case of doubtful identification. Resistance was verified by checking the inhibition zone diameter of under 20 mm on 30 microg cefoxitin disks. RESULTS: Bio-Rad Laboratories, Becton Dickinson and bioMérieux media read at 24 or 24/48 hours have a respective sensitivity of 91, 71 and 85/92%. The specificity of these media is 99, 100 and 99/93%. CONCLUSION: These media proved to be new powerful and rapid tools used in screening for MRSA detection. MRSASelect is one of the most effective media showing high sensitivity and specificity and the easiest interpretation. chromID MRSA exhibits similar performance but needs more time to be as effective as Bio-Rad media while CHROMagar MRSA isn't enough efficient with its slightly lack of sensitivity to perform a reliable screening.     

Evaluation of two new chromogenic media, CHROMagar MRSA and S. aureus ID, for identifying Staphylococcus aureus and screening methicillin-resistant S. aureus

Thirty-nine methicillin-resistant Staphylococcus aureus (MRSA) isolates with diverse genetic backgrounds and two reference strains were correctly identified as S. aureus on CHROMagar MRSA and S. aureus ID media. Growth inhibition on CHROMagar MRSA was noted. A combination of cefoxitin disk and S. aureus ID was found suitable for rapid MRSA screening.     

Performance of MRSA ID, a new chromogenic medium for detection of methicillin-resistant Staphylococcus aureus

MRSA ID was evaluated for its ability to identify methicillin-resistant Staphylococcus aureus. A well-defined collection of staphylococci was used (n = 998). The sensitivity after 24 h was 96.4%, increasing to 98.8% after 48 h. The specificity was 98.2% after 24 h and decreased to 89.7% after 48 h.     

Strain differentiation in methicillin-resistant Staphylococcus aureus

Three different systems were used to test 236 isolates of methicillin-resistant Staphylococcus aureus in an attempt to ascertain if more than one strain is responsible for the current problem of cross-infection by this organism in N.S.W. hospitals. The biochemical tests used were of little assistance. Phage typing, using the Basic International Set of typing phages at 100 x routine test dilution (RTD), provided evidence of the presence of several different strains. Phage type 83A/85/95/90/88 was the typing pattern of the predominant strain and the nest most frequent group was not typable. These results were often difficult to read. Five new phages were therefore isolated and found to be valuable as they produced easily identifiable patterns at RTD.     

Gentamicin resistance in methicillin-resistant Staphylococcus aureus

Gentamicin resistance has been studied in methicillin-resistant Staphylococcus aureus (MRSA) strains, from Royal Melbourne Hospital (RMH) and Sydney. Gentamicin resistance was transferred in mixed cultures to a plasmid free strain, and the determinants were examined. The Sydney strain had high level resistance to gentamicin, tobramycin, kanamycin and neomycin which was carried on a c.34 megadalton plasmid. The gentamicin resistant RMH isolates all had a determinant which conferred low level resistance to gentamicin, tobramycin and kanamycin and appeared to be chromosomal in one isolate, on a plasmid of c.28.5 megadaltons in another and on a plasmid of c.18 megadaltons in the other isolates. It is suggested that a gentamicin resistance transposon is being transferred in the MRSA at RMH.     

Evaluation of three chromogenic media (MRSA-ID, MRSA-Select and CHROMagar MRSA) and ORSAB for surveillance cultures of methicillin-resistant Staphylococcus aureus

Screening specimens were homogenised in saline 0.9% w/v before either direct inoculation or following enrichment in broth on three chromogenic media (MRSA-ID, CHROMagar MRSA and MRSA Select) and ORSAB medium for the detection of methicillin-resistant Staphylococcus aureus (MRSA). In total, 102 of 466 specimens yielded MRSA on at least one medium. After incubation for 16-18 h, the sensitivity was 51%, 59%, 47% and 65% on MRSA-ID, CHROMagar MRSA, ORSAB and MRSA Select, respectively, compared with 82%, 75%, 67% and 80%, respectively, after 42 h, and 93%, 95%, 79% and not tested, respectively, following broth enrichment. There were significantly more MRSA colonies on MRSA-Select after 16-18 h than on ORSAB or MRSA ID (p 0.001 and 0.0022, respectively), whereas there were more MRSA colonies after 42 h on MRSA-ID and MRSA-Select than on ORSAB (p 0.0004 and 0.012, respectively). The specificity of the media for identifying MRSA based on the colour of colonies after incubation for 16-18 h was 100%, 99%, 99% and 100%, respectively, compared with 98%, 97%, 98% and 98%, respectively, after 42 h, and 100%, 99%, 100% and not tested, respectively, following broth enrichment. The speed of detection (mean time to report a positive result) was 1.65, 1.72, 2.31 and 1.35 days, respectively. For each of the three media tested following enrichment, the use of an enrichment broth increased the detection rate of MRSA by 16-24%.     

Screening media for detection of methicillin-resistant Staphylococcus aureus from non-sterile body sites

Four hundred and forty clinical isolates were tested on Baird-Parker and Vogel-Johnson agars with 6 g/ml of oxacillin, to determine their growth characteristics on these potential screening media for methicillin-resistant Staphylococcus aureus (MRSA). While both media perfomed well individually, a combination biplate with each medium may be the most useful in screening patients for MRSA from normally non-sterile sites.     

Evaluation of a new chromogenic medium, MRSA select, for detection of methicillin-resistant Staphylococcus aureus

We compared MRSA Select to mannitol-salt agar with 8 microg/ml cefoxitin for the detection of methicillin-resistant Staphylococcus aureus (MRSA) from 6,199 clinical samples submitted for MRSA screening. The sensitivities and specificities of MRSA Select and mannitol-salt agar with cefoxitin were 98% and 92% versus 90% and 78%, respectively (P<0.0001). Most (96%) MRSA were detected after overnight incubation using MRSA Select.     

Community-acquired methicillin-resistant Staphylococcus aureus in Taiwan.

Staphylococcus aureus is a major cause of infections in both hospitals and communities, and is exhibiting increasing resistance to methicillin (methicillin-resistant S. aureus, MRSA) and related beta-lactams. MRSA is usually considered a nosocomial pathogen, but increasingly it is acquired in the community. In Taiwan, MRSA was colonized in a substantial proportion of healthy children and accounted for 25% to 75% of childhood community-acquired (CA) S. aureus infections. From the preliminary data, the isolates of sequence type (ST) 59 by multilocus sequence typing method appeared to be the major clone of CA-MRSA in northern Taiwan. Compared with those reported from the US and other countries, CA-MRSA isolates in Taiwan did not always harbor type IV staphylococcal cassette chromosome (SCCmec) and were resistant to multiple non-beta-lactam antibiotics, including clindamycin and macrolides. Molecular evidence suggested transmission of the community strain of MRSA into the hospital setting, and that the community strain had became a health care-associated pathogen. The treatment of putative CA S. aureus infection should be stratified according to the severity and the disease entity.     

Mechanism of beta-lactam-resistance in methicillin-resistant Staphylococcus aureus

The role of penicillin-binding protein (PBP) 2' in the expression of beta-lactam-resistance was investigated using methicillin-resistant Staphylococcus aureus (MRSA) strains with different level of resistance. Both high- and moderate-level MRSA produced very similar PBP 2' with low affinities for beta-lactam antibiotics. Affinities of antibiotics for PBP 2' (I50, concentration which inhibits [14C] benzylpenicillin-binding by 50%) correlated well with their antibacterial activities (MIC) in a high-level MRSA, but did not in a moderate-level MRSA. High-level MRSA contained a larger amount of PBP 2' than moderate-level MRSA, and the amount of PBP 2' decreased by increasing the temperature of the culture; the extent of decrease was larger in a strain which was sensitive at 37 degrees C than a strain which exerted relatively high level resistance even at 40 degrees C. A cephamycin-resistant, methicillin-sensitive strain began to synthesize PBP 2' by adding cephamycin-type antibiotics to the medium and consequently acquired resistance to methicillin. Latent MRSA producing no PBP 2' generated clones which produced PBP 2' constitutively and were highly resistant to all beta-lactams. These results suggest that the presence of PBP 2' is critical for the expression of beta-lactam-resistance in MRSA and the degree of the resistance depends mainly on the amount of PBP 2' which differs from strain to strain and is influenced by environments such as temperature and the presence of inducer.     

Detection of methicillin-resistant Staphylococcus aureus in clinical specimens from cystic fibrosis patients by use of chromogenic selective agar

We evaluated the use of a chromogenic selective medium (MRSA ID) as a useful tool for the detection of methicillin-resistant Staphylococcus aureus (MRSA) in cystic fibrosis (CF) patient samples. Fifty-four MRSA isolates were detected by MRSA ID, while only 24/54 (44%) (odds ratio [OR], 2.79; 95% confidence interval [CI], 1.63 to 4.76) were detected by conventional methods. A chromogenic selective medium for MRSA detection may improve its surveillance in CF patients.