Evaluation of a new selective medium for methicillin-resistant Staphyloccocus aureus

The aims of this study were to evaluate the performance of a new medium, desferrioxamine oxacillin tellurite egg-yolk mannitol salt agar (DOTEMSA) in detecting methicillin-resistant Staphylococcus aureus (MRSA) and then to compare this medium against the Public Health Laboratory Service (PHLS) recommendation of mannitol salt agar (Oxoid) with oxacillin (OMSA) and Baird-Parker medium with ciprofloxacin (BPC) for the isolation of MRSA. The individual selective agents contained in DOTEMSA were tested against isolates of coagulase-negative staphylococci (CNS) and the medium with all constituents was challenged with various bacteria. Routine screening specimens were plated out on OMSA, BPC and DOTEMSA and the plates were incubated and examined at 24 and 48 h. Tellurite, desferrioxamine and oxacillin each inhibited the majority of CNS isolates; only three (of 103) grew in the presence of all three agents. Sixty-two of 63 isolates of MRSA grew on DOTEMSA and 59 produced lipase. Most other bacteria were inhibited. In all, 184 MRSA isolates were isolated from 540 screening specimens. The sensitivity of OMSA, BPC and DOTEMSA was 42%, 81% and 51% at 24 h, and 60%, 89% and 89% at 48 h. At 48 h, the combination of BPC and DOTEMSA detected 99% of MRSA isolates. Seventy, 49 and one non-MRSA isolates needed investigation for each of the three media respectively. A proposed strategy for MRSA screening would use BPC and DOTEMSA, examining BPC at 24 h and both media at 48 h. Provisional reports could then be issued at 24 h on the basis of rapid agglutination tests to confirm isolates as S. aureus from BPC and at 48 h on the basis of typical colonies from DOTEMSA.     

Comparison of methods for the isolation of methicillin resistant Staphylococcus aureus.

The control of methicillin resistant Staphylococcus aureus (MRSA) relies on the rapid and sensitive detection of carriage. The roles of an enrichment broth, duration of incubation, and Baird-Parker medium containing ciprofloxacin (BPC) were evaluated in comparison with standard media in a centre where the prevalence of ciprofloxacin resistance among MRSA is over 98%. Screening swabs from 402 sites were plated onto BPC, mannitol salt agar (MSA), and MSA with methicillin (MMSA). The swabs were enriched in Tryptone-T broth with 6% salt for 24 hours and the broths subcultured onto BPC, MSA, and MMSA. MRSA was isolated from 134 swabs. Significantly more isolates were obtained by incubating culture plates for 42 hours rather than 18 hours, by the use of broth enrichment, and by addition of methicillin or ciprofloxacin to media. BPC was the most sensitive medium (107 isolates (80%) by direct culture at 42 hours), grew the fewest contaminants, and allowed provisional reporting of 73% of isolates at 18 hours by colonial appearance and use of Staphaurex Plus rapid latex reagent. This may allow the introduction of infection control measures a day earlier than when other established methods are used.     

Screening for methicillin resistance in Staphylococcus aureus and coagulase-negative staphylococci: an evaluation of three selective media and Mastalex-MRSA latex agglutination

Laboratory confirmation of MRSA is important for the implementation of infection control; conventional screening culture methods take up to five days for confirmation. The purpose of this study is to ascertain the efficiency of three selective media for growth of methicillin-resistant Staphylococcus aureus (MRSA) before and after enrichment in salt broth, and to evaluate the Mastalex-MRSA latex agglutination kit for detection of methicillin resistance. Screening swabs were collected from 63 patients, yielding 125 S. aureus isolates and 40 coagulase-negative staphylococcus (CNS) isolates. Selective media used were mannitol salt agar (MSA), Baird-Parker agar with ciprofloxacin (BPC) and bromocresol purple (BCPA). Polymerase chain reaction (PCR) for mecA gene detection was used as the reference standard for evaluation of the Mastalex-MRSA assay, which was also evaluated on colonies of S. aureus from the selective media. No significant difference was found in efficiency of MRSA isolation among the selective media. Pre-enrichment in the salt broth did not enhance isolation of MRSA. Methicillin-sensitive S. aureus and CNS were significantly inhibited in all selective media (P<0.05). Only BPC significantly selected out methicillin-resistant CNS (P<0.01). Mastalex-MRSA was 97% specific and sensitive for the detection of MRSA. It was 65% sensitive and 100% specific in detecting methicillin resistance in CNS. In conclusion, all selective media performed equally well (MRSA isolation rate of approximately 80%). Mastalex-MRSA provided rapid and reliable detection of MRSA from selective media, reducing the time required for confirmation of this organism.     

Catheter-associated fungemia caused by Fusarium chlamydosporum in a patient with lymphocytic lymphoma.

We evaluated the use of mannitol salt agar with oxacillin for use as a primary screening medium for the simultaneous detection and identification of methicillin-resistant Staphylococcus aureus in clinical surveillance specimens. Oxacillin agar dilution susceptibility tests with mannitol salt agar and Mueller-Hinton agar were performed in parallel with disk-agar diffusion testing on 95 oxacillin-susceptible and 105 oxacillin-resistant S. aureus stock isolates. MICs were found to be comparable, showing distinct separation of susceptible and resistant isolates into two groups with MICs of less than or equal to 2 and greater than or equal to 32 micrograms/ml, respectively. In accord with these findings, 4 micrograms of oxacillin per ml was selected for use in the screening medium. For performance evaluation, mannitol salt agar with 4 micrograms of oxacillin per ml was compared with mannitol salt agar without oxacillin by performing parallel screening tests on 153 clinical surveillance specimens. For detection of methicillin-resistant S. aureus, mannitol salt agar with 4 micrograms of oxacillin per ml was as sensitive as mannitol salt agar without oxacillin and required significantly fewer confirmatory tests. For primary identification of methicillin-resistant S. aureus, mannitol salt agar with 4 micrograms of oxacillin per ml was 6.4% false-positive and 1.1% false-negative, with a 93.6% positive predictive value. These findings indicate that mannitol salt agar with 4 micrograms of oxacillin per ml can be used as a reliable and cost-effective screening medium for the simultaneous detection and identification of methicillin-resistant S. aureus in clinical surveillance specimens.     

Mannitol salt agar-cefoxitin combination as a screening medium for methicillin-resistant Staphylococcus aureus

In disk diffusion tests, cefoxitin is now considered a better indicator than oxacillin for the presence of the mecA gene in Staphylococcus aureus. A logical extension of this work is the incorporation of cefoxitin into media selective for methicillin-resistant Staphylococcus aureus (MRSA). This paper describes the development and subsequent testing of mannitol salt agar containing 4 mg/liter cefoxitin with a unique collection of well-characterized MRSA strains, including low-level methicillin-resistant strains and an equal number of known mecA-negative S. aureus strains. The agar supported the growth of 96.6% of the mecA-positive strains in the collection and inhibited the growth of 100% of the mecA-negative strains. These results suggest that selective media based on cefoxitin are superior to those based on oxacillin for the detection of MRSA.     

Methicillin-resistant Staphylococcus aureus: evaluation of detection techniques on laboratory-passaged organisms

Strains of methicillin-resistant Staphylococcus aureus (MRSA) are a major cause of nosocomial infection. This has led to the widespread introduction of screening techniques to identify patients or staff colonised with the organism and prevent the spread of MRSA in the hospital environment. The aim of laboratory techniques for the detection of MRSA screening is the rapid isolation and identification of MRSA, enabling timely implementation of appropriate infection control measures. This study compares commercially available products used by the majority of laboratories during routine screening of samples for MRSA. Several selective media are tested, including mannitol salt agar, Mueller Hinton agar, milk agar and blood agars. The accuracy of combining rapid agglutination from selective media for identification of MRSA is also determined, using six commercially produced kits. Owing to debate concerning the use of enrichment broth, in addition to primary isolation on solid media, four enrichment broths are tested and compared to direct culture techniques. Using a selection of laboratory-passaged MRSA phage types, results demonstrated that mannitol salt agar containing 75 g/L NaCl, without the addition of oxacillin, recovered all 50 MRSA strains and produced the highest isolation rate. Robertson's cooked-meat broth, with 7.5% NaCl, proved the most effective enrichment medium and was more sensitive than direct culture by 3%. Pastorex Staph-plus proved to be the most efficient rapid agglutination kit when testing MRSA colonies removed directly from selective agars.     

SCREENING FOR METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS CARRIERS AMONG PATIENTS AND HEALTH CARE WORKERS OF A TERTIARY CARE HOSPITAL IN SOUTH INDIA

A total of 200 subjects were screened for carriage of methicillin-resistant Staphylococcus aureus (MRSA) at different sites using oxacillin blood agar and mannitol salt agar with oxacillin. Overall carriage rate was 8.5%, with the highest rate in inpatients (15.6%) while the lowest was seen in health care workers (1.8%). The commonest site of colonization was the anterior nares. Oxacillin blood agar was found to be superior to mannitol salt agar with oxacillin for the isolation of MRSA. Male sex and prolonged hospital stay were found to be the major risk factors for MRSA colonization.     

Comparison of different techniques for the detection of heterogeneous resistance to methicillin in Staphylococcus aureus]

The methods for detection of methicillin resistant S. aureus (MRSA) can fail to detect resistance because phenotypic expression is often heterogeneous (40% of strains). Seventy four strains of S. aureus [4 methicillin susceptible strains, 10 homogeneous MRSA (Ro) and 60 heterogeneous MRSA (Rh)] were isolated from different french hospitals in Paris. These strains were tested by different methods: oxacillin screen plate with 6 micrograms/ml oxacillin and 4% NaCl, agar diffusion method with 5 micrograms oxacillin disk tested either at 30 degrees C on Mueller-Hinton medium or at 37 degrees C on Mueller-Hinton plus 5% NaCl, BBL Crystal MRSA ID system tested with two inocula (0.5 and 1 McFarland equivalent bacterial suspension) at 37 degrees C for 4 h and 5 h. Dot-blot hybridization was performed under stringent condition with the mecA probe. The accuracy of the different methods for the detection of methicillin resistance is equivalent, except for the BBL crystal system with a 0.5 McFarland inoculum wich detects the resistance with an accuracy of 86% for Ro strains and 69% for Rh strains. In other respects, there was a close correlation with the detection of the phenotypic resistance and the presence of mecA gene. So this study demonstrates that these various methods can be used for the detection of methicillin resistant S. aureus. For a rapid detection (below 5 h) the BBL crystal system with a 1 McFarland inoculum can be used; the agar diffusion method remains a good method provided some conditions (inoculum, incubation temperature, addition of salt, incubation period); the oxicillin screnn plate is a very attractive method for it is easy and reliable.     

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.     

Application of lipovitellin-salt-mannitol agar for screening, isolation, and presumptive identification of Staphylococcus aureus in a teaching hospital.

Lipovitellin-salt-mannitol (LSM) plate medium was examined for its ability to directly isolate, recover, and presumptively identify Staphylococcus aureus from 418 clinical specimens. The criteria for medium evaluation included colony morphology reactions, selectivity, and ease of isolation. For 298 specimens used for screening, LSM agar medium was compared with the other conventional media used, mannitol salt agar (MSA), 5% horse blood agar (HBA), and phenolphthalein phosphate agar (PPA), to detect and recover S. aureus and methicillin-resistant S. aureus. The results indicated that LSM agar is more effective than MSA, HBA, or PPA for the recovery and isolation of S. aureus and methicillin-resistant S. aureus. On a replicator multipoint inoculation system, we compared the reactions on LSM agar, MSA, and DNase agar of 227 different strains of staphylococci, which included 178 different strains of S. aureus and 49 different strains of coagulase-negative staphylococci isolated from clinical specimens. By using the lipovitellin precipitation activity and mannitol fermentation characteristics, LSM agar gave a 100% correlation in presumptively identifying S. aureus. LSM agar may be an alternative plate medium for large hospital extensive screening for the detection and isolation of S. aureus.     

Latex agglutination for rapid identification of methicillin-resistantStaphylococcus aureus recovered from selective media

The accuracy of combining latex agglutination with selective media for the identification of methicillin-resistantStaphylococcus aureus (MRSA) was determined. Test strains were identified by latex agglutination on blood agar, the heat-stable thermonuclease test and broth microdilution MICs of oxacillin and included 97 MRSA, 56 methicillin-susceptibleStaphylococcus aureus, 52 methicillin resistant, and 49 methicillin-susceptibleStaphylococcus species. Isolates were grown on trypticase-soy agar with 5 % sheep red blood cells (TSAB), Mueller-Hinton agar (MHA), mannitol-salt agar (MSA), and four media designed for the selective growth of MRSA: TSAB with clindamycin and gentamicin, MHA with oxacillin, MSA with oxacillin, and lipovitellin-salt-mannitol agar (LVSM) with 1 µg oxacillin disks applied. The mean sensitivity, specificity, and positive predictive value for the combination of latex agglutination with selective media for the identification of MRSA was 96 %, 99 % and 98 % respectively.     

Comparison of screening methods to identify methicillin-resistantStaphylococcus aureus

Screening methods to identify methicillin-resistantStaphylococcus aureus (MRSA) were compared using 96 isolates representing 17 distinct clones. The sensitivity of four commercial agglutination tests was determined in comparison to the tube coagulation test, and the results related to the presence of the coagulase gene. The broth screening test, agar dilution test and disc diffusion test were carried out, and the results related to the presence of themecA gene. Mannitol salt agar and Iso-Sensitest agar with varying salt supplements were used. All agglutination tests had high rates of detection ofStaphylococcus aureus (95.8–99.0%). Resistance in mecA gene-positiveStaphylococcus aureus isolates was correctly detected by the oxacillin broth test, the agar dilution test and the disc diffusion test on mannitol salt agar, whereas on Iso-Sensitest agar detection rates were lower (between 68.5% and 94.4%, depending on the salt supplement). Incubation of the Iso-Sensitest plates for 48 hours significantly improved the rate of detection of resistance, but increased the major error rate up to 71.4%.MecA genepositiveStaphylococcus aureus isolates not detected by the disc diffusion test on Iso-Sensitest agar had significantly lower oxacillin minimal inhibitory concentration values and were significantly less resistant to a variety of antibiotics. Thus, mannitol salt agar might be a suitable medium for use in the disc diffusion and agar dilution test to detect resistance to oxacillin inStaphylococcus aureus.     

Comparison of protocols for surveillance of methicillin-resistant Staphylococcus aureus (MRSA): medical staff vs ICU patients

To compare the sensitivity of various protocols for methicillin-resistant Staphylococcus aureus (MRSA) surveillance, active surveillance for detecting MRSA nasal colonization was performed on 97 members of the medical staff and 218 patients in the Intensive Care Unit (ICU) of a university hospital. Duplicate nasal swabs were collected from each participant. One was plated directly on a blood agar plate (D-BAP) and observed at 24 and 48 hr. Another was incubated overnight in tryptic soy broth (TSB) with 6.5% NaCl, and subcultured on both BAP (B-BAP) and mannitol salt agar with 4 mg/L of oxacillin (B-MSAOXA). The MRSA colonization rate was similar in the medical staff and patient samples (16.5% vs 11.9%, p = 0.285). Among the medical staff members, the sensitivity of MRSA detection was the same (93.8%) in D-BAP and B-BAP. In the ICU patients, which are a high-risk group, the sensitivity of MRSA detection was improved by adding a pre-enrichment step (73.1% on D-BAP vs 96.2% on B-BAP). The simple direct plating protocol was sufficiently sensitive for the medical staff members, but pre-enrichment was an essential step to increase detection of MRSA in the ICU patients.     

Comparison of culture screening methods for detection of nasal carriage of methicillin-resistant Staphylococcus aureus: a prospective study comparing 32 methods

Screening for carriage of methicillin-resistant Staphylococcus aureus (MRSA) is fundamental to modern-day nosocomial infection control, both for epidemiologic investigation and day-to-day decisions on barrier isolation. Numerous microbiologic techniques have been advocated for screening for nasal carriage of MRSA, including the use of charcoal rather than rayon swabs, preincubation of swabs in Stuart's medium, preincubation of swabs in salt-containing trypticase soy broth (TSB), use of mannitol-salt agar (MSA), use of MSA containing oxacillin (MSA(Ox)), use of Mueller-Hinton agar containing oxacillin (MHA(Ox)), and the use of MSA containing lipovitellin with an oxacillin disk (MSAL(Ox)). We report a prospective clinical trial undertaken to test all of these methods concurrently. Patients at high risk for MRSA carriage were screened with eight consecutive nasal swabs (four standard rayon, four charcoal-coated rayon), which were processed by primary plating on MSA, MSA(Ox), MHA(Ox), and MSAL(Ox); Stuart's preincubation for 72 h followed by plating on the solid media; overnight enrichment in salt-containing TSB followed by plating; and Stuart's preincubation for 72 h followed by overnight enrichment in TSB and plating. All of the above methods were repeated with charcoal swabs. Each patient was screened by 32 culture methods. Forty-three (42%) of 102 patients studied were positive for MRSA by one or more methods. Among the four media evaluated with direct plating, MSAL(Ox) was 11 to 25% more sensitive for detecting MRSA (MSAL(Ox) versus MSA(Ox) or MHA(Ox) or MSA, each P < 0.01). Preincubation in Stuart's medium for 72 h did not enhance recovery of MRSA. Enrichment in salt-containing TSB further increased yield 9%. MSAL(Ox) also showed the best specificity, 93%. Charcoal swabs showed no advantage over standard rayon swabs. Our results suggest that the highest yield will be achieved by using standard rayon swabs that are enriched overnight in TSB with inoculation onto MSAL(Ox) medium. Direct inoculation of swabs onto MSAL(Ox) allows detection of 90% of MRSA carriers.     

Multicenter evaluation of BBL CHROMagar MRSA medium for direct detection of methicillin-resistant Staphylococcus aureus from surveillance cultures of the anterior nares

Active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) is among the strategies recommended by the Society for Healthcare Epidemiology of America for control of nosocomial MRSA infections. Infection control and laboratory personnel desire rapid, sensitive, and inexpensive methods to enhance surveillance activities. A multicenter study was performed to evaluate a new selective and differential chromogenic medium, BBL CHROMagar MRSA (C-MRSA) medium (BD Diagnostics, Sparks, MD), which enables recovery and concomitant identification of MRSA strains directly from nasal swab specimens taken from the anterior nares. Specimens were inoculated to C-MRSA and Trypticase soy agar with 5% sheep blood agar (TSA II, BD Diagnostics). Mauve colonies on C-MRSA at 24 h and 48 h and suspicious colonies on TSA II were confirmed as Staphylococcus aureus by Gram stain morphology and a coagulase test. In addition, the results of C-MRSA were compared to results of susceptibility testing (five different methods) of S. aureus strains isolated on TSA II. A total of 2,015 specimens were inoculated to C-MRSA and TSA II. Three hundred fifty-four S. aureus isolates were recovered; 208 (59%) were oxacillin (methicillin) susceptible and 146 (41%) were oxacillin resistant (MRSA). On C-MRSA, 139/146 or 95.2% of MRSA isolates were recovered, whereas recovery on TSA II was 86.9% (127/146) (P = 0.0027). The overall specificity of C-MRSA was 99.7%. When C-MRSA was compared to each susceptibility testing method, the sensitivity and specificity, respectively, were as follows: oxacillin MIC by broth microdilution, 94.4% and 96.7%; oxacillin screen agar, 94.3% and 96.7%; PBP2' latex agglutination, 93.7% and 98.5%; cefoxitin disk diffusion, 95.0% and 98.1%; and mecA PCR, 95.1% and 98.1%. In this study, C-MRSA was superior to TSA II for recovery of MRSA from surveillance specimens obtained from the anterior nares and was comparable to conventional, rapid, and molecular susceptibility methods for the identification of MRSA isolates.     

Evaluation of a novel medium for screening specimens from hospitalized patients to detect methicillin-resistant Staphylococcus aureus

A novel medium, Oxacillin Resistant Screening Agar (ORSA) medium, was evaluated for the screening of specimens for methicillin-resistant Staphylococcus aureus (MRSA) in the hospital setting. Screening swabs (swabs of the nose, throat, perineum, and infected sites) were inoculated onto the new ORSA medium and into an enrichment broth (Muller-Hinton broth supplemented with NaCl and oxacillin). After 24 h of incubation, the enrichment broth was subcultured onto one ORSA plate and one lipovitellin Chapman salt agar plate. The sensitivities for the detection of MRSA were calculated for each medium alone and for the media in combination. A low sensitivity (74%) was obtained when ORSA medium was used alone as a primary culture, whereas the sensitivity was 88% when a single selective enrichment broth was used. Among the 414 blue colonies observed on ORSA plates, only 47% were found to be MRSA, 40% were coagulase-negative staphylococci, 7% were Enterococcus species, and 2% were methicillin-sensitive S. aureus. The optimal incubation time for the ORSA plates was evaluated. On primary culture, 38% of the blue MRSA colonies were visible only after 48 h of incubation (no blue colonies were not seen after 24 h of incubation), whereas 94% of the colonies were already visible at 24 h when ORSA plates were used for subcultures. In conclusion, the advantage of the novel ORSA medium is the ease of recognition of mannitol-fermenting bacteria, but further identification tests are needed to confirm the identification of S. aureus. An enrichment broth is still needed to ensure a good sensitivity for the recovery of MRSA, and an incubation time of 48 h is required for primary culture on ORSA medium.     

Detection of low-level oxacillin resistance in mecA-positive Staphylococcus aureus

Detection of low-level oxacillin-resistant Staphylococcus aureus is a problem that needs special attention, particularly in relation to methicillin-resistant S. aureus (MRSA) strains in the community that belong to clonal lineage ST80. This study compared different phenotypic methods for the detection of 74 low-level oxacillin-resistant S. aureus strains (oxacillin MIC or=2 mg/L) and 117 methicillin-susceptible S. aureus strains. Determination of microbroth dilution MICs for oxacillin was wholly unsatisfactory, and gave a limited specificity for cefoxitin. The sensitivity of disk-diffusion performed according to CLSI recommendations was 92% with an oxacillin 1-microg disk, and 96% with a cefoxitin 30-microg disk; use of a 10-microg cefoxitin disk and a semi-confluent inoculum (breakpoint for resistance <18 mm zone diameter) gave a sensitivity of 97%. When disk-diffusion was performed on IsoSensitest agar with a zone diameter breakpoint for resistance of <22 mm (as recommended by the Swedish Reference Group for Antibiotics), the sensitivity was 95%.     

Screening method for recovery of methicillin-resistant Staphylococcus aureus from primary plates.

A study designed to screen for the presence of methicillin-resistant Staphylococcus aureus from primary plates was conducted from 1 January to 1 September 1985 in a small community hospital. The screening method used a plate of lipovitellin salt mannitol agar and a 4-microgram oxacillin disk incubated at 30 degrees C. Growth of yellow colonies, typical of S. aureus, around the disk without a zone of inhibition was called presumptive methicillin-resistant S. aureus. All susceptibilities were confirmed by using the National Committee for Clinical Laboratory Standards macrodilution technique. Of 224 cultures containing S. aureus, 118 (53%) were positive for methicillin-resistant S. aureus isolates. Of these 118, 111 (94%) were correctly identified from the primary plates as methicillin-resistant S. aureus. Of the 224 isolates, 14 could not be categorized from the primary plates as methicillin-resistant S. aureus due to the small amounts of S. aureus recovered.     

Prospective comparison of a new chromogenic medium, MRSASelect, to CHROMagar MRSA and mannitol-salt medium supplemented with oxacillin or cefoxitin for detection of methicillin-resistant Staphylococcus aureus

MRSASelect agar was compared to CHROMagar, mannitol-salt agar with oxacillin, and mannitol-salt agar with cefoxitin (MSA-CFOX) for the isolation of methicillin-resistant Staphylococcus aureus (MRSA). The sensitivities and specificities were 97.3% and 99.8%, 82.9% and 99.1%, 80.2% and 79%, and 99.1% and 84.8%, respectively. MSA-CFOX and MRSASelect had a high sensitivity. MRSASelect, however, was more specific and proved to be a more reliable and rapid medium for the detection of MRSA.     

Rapid Slide Latex Agglutination Test for Detection of Methicillin Resistance in Staphylococcus aureus

The MRSA screen test (Denka Seiken Co., Ltd.), a commercially available, rapid (20-min) slide latex agglutination test for the determination of methicillin resistance by detection of PBP 2a in Staphylococcus aureus, was compared with the oxacillin agar screen test and PCR detection of the mecA gene. A total of 563 S. aureus isolates were tested. Two hundred ninety-six of the isolates were methicillin-susceptible isolates from cultures of blood from consecutive patients. Also, 267 methicillin-resistant isolates that comprised 248 different phage types were tested. Methicillin resistance was defined as the presence of the mecA gene. Of the 267 mecA gene-positive isolates, 263 were positive by the MRSA screen test (sensitivity, 98.5%), and all the mecA-gene negative strains were negative by the MRSA screen test (specificity, 100%). The oxacillin agar screen test detected methicillin resistance in 250 of the mecA gene-positive isolates (sensitivity, 93.6%). The sensitivity of the MRSA screen test was statistically significantly higher than the sensitivity of the oxacillin agar screen test (P < 0. 05). The MRSA screen test is a highly sensitive and specific test for the detection of methicillin resistance. Also, it offers results within half an hour and is easy to perform, which makes this test a valuable tool in the ongoing battle against methicillin-resistant S. aureus.