Unusual susceptibility of methicillin-resistant Staphylococcus aureus to erythromycin, clindamycin, gentamicin, and tetracycline at 30 degrees C but not at 35 degrees C.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important pathogen in hospital environments, and optimal detection of MRSA requires nonroutine methods in clinical microbiology laboratories. One such method is an incubation temperature of 30 degrees C in contrast to the more commonly used temperature of 35 degrees C. To determine the percentage of MRSA isolates that would be missed if only one temperature were used, we evaluated methicillin resistance and susceptibility of 2,397 S. aureus isolates by agar dilution at 30 and 35 degrees C. Of the clinical isolates, 93% showed matching MICs of methicillin at both temperatures. Another 6.8% (162) showed different MICs at 30 and 35 degrees C, with 60 of the 162 isolates (2.5% of all isolates studied) being resistant to methicillin at 30 degrees C but susceptible at 35 degrees C (temperature-discrepant MRSA). MICs of other antimicrobial agents, measured at 35 degrees C, revealed an unusual pattern of susceptibility of these temperature-discrepant MRSA isolates. In contrast to previously reported resistance of MRSA at 35 degrees C to erythromycin, clindamycin, gentamicin, and tetracycline, the temperature-discrepant MRSA isolates were susceptible to these agents. This resistance pattern may be of value in identifying questionable MRSA isolates when only one incubation temperature is used.     

Validation of multiplex PCR strategy for simultaneous detection and identification of methicillin resistant Staphylococcus aureus

Multiplex polymerase chain reaction (PCR) strategy is described for rapid identification of clinically relevant methicillin resistant Staphylococcus aureus (MRSA) that targets mecA and coagulase genes. In this study, 150 staphylococcal clinical isolates were used that included 40 isolates of MRSA, 55 isolates of methicillin susceptible S. aureus (MSSA), 44 isolates of methicillin susceptible coagulase negative Staphylococcus spp. (MS-CoNS) and 11 isolates of methicillin resistant coagulase negative Staphylococcus spp. (MR-CoNS). Out of 55 S. aureus strains, three strains demonstrated mecA gene, which appeared to be oxacillin sensitive by disc diffusion. When (MS-CoNS) were evaluated, 10 isolates classified as oxacillin sensitive phenotypically, yielded positive results in PCR method. The results for mecA detection by PCR were more consistent with disk susceptibility tests in case of MRSA (100%) and MSSA (95%) isolates. In contrast to above results with MRSA and MSSA, mecA detection by PCR in MS-CoNS showed less correlation with disk susceptibility tests (77%). The results for coag detection by PCR were consistent with phenotypic tests in all isolates.     

Improved and rapid detection of methicillin-resistant Staphylococcus aureus nasal carriage using selective broth and multiplex PCR

To improve efficiency in detecting nasal methicillin-resistant Staphylococcus aureus (MRSA), we evaluated a multiplex PCR using pre-enrichment of the specimen in selective broths, and compared it with detection performed by routine tests in hospital laboratories. Nasal swab specimens from 311 patients were inoculated onto mannitol-salt agar (MSA) at the hospital laboratories and in two Mueller-Hinton broths with 7% NaCl containing oxacillin at concentrations of 2 and 4 micro g/ml. Isolates on MSA were identified as MRSA by classical laboratory tests (coagulase and oxacillin disk diffusion tests). Oxacillin broth cultures were subcultured on blood agar and MRSA isolates were identified by coagulase and susceptibility tests, including agar dilution and the oxacillin-screening method (gold standard method). Simultaneously, multiplex-PCR was performed from the selective broths to detect S. aureus species-specific and mecA gene segments (OxMPCR method). Thirty-two S. aureus isolates were recovered: 29 (90.6%) were MRSA strains and 3 (9.4%) were oxacillin-susceptible isolates. Twenty-eight (96.5%) MRSA isolates were detected by OxMPCR, while 17 (58.6%) were identified by routine tests (P=0.002). This new method for detection of MRSA nasal carriers showed higher sensitivity and led to faster reporting--i.e., within 24 h--of results.     

Nationwide surveillance for Staphylococcus aureus with reduced susceptibility to vancomycin in Korea

Methicillin-resistant Staphylococcus aureus (MRSA) accounts for more than 70% of S. aureus isolates from tertiary hospitals in Korea. Clinical isolates of S. aureus were collected from eight provincial, university-affiliated hospitals during the period from June 1999 to January 2001 for nationwide surveillance. All isolates were screened for reduced susceptibility to vancomycin by using brain heart infusion agar containing 4 micro g of vancomycin per milliliter. Population analysis and the determination of the MIC of vancomycin were done for the isolates which grew on the screening agar plates. Of 682 total isolates, MRSA accounted for 64% (439 of 682). Of 27 (4%) isolates that grew on the screening agar plates, none showed the heteroresistance phenotype. No strains with reduced susceptibility to vancomycin were identified.     

Molecular pattern and antimicrobial susceptibility of methicillin-resistant Staphylococcus aureus isolates at a teaching hospital in Northern Taiwan.

BACKGROUND AND PURPOSE: Methicillin-resistant Staphylococcus aureus (MRSA) infection has progressively increased worldwide. Knowledge of the specific epidemiological pattern of isolates at individual hospitals is important. METHODS: MRSA bacteremia was diagnosed in a total of 68 patients from January 2002 through December 2003, stratified for drug susceptibility and molecular pattern (staphylococcal cassette chromosome mec element [SCCmec] typing and genotypes). RESULTS: SCCmec-A-positive isolates were found on polymerase chain reaction in 58 patients. The most frequent SCCmec types were III (40 cases) of which less than 5% were susceptible to other beta-lactam antibiotics and most were health care-associated, followed by SCCmec type IV (15 cases), that were demonstrated to be community-acquired. SCCmec type IV MRSA isolates were more likely to be susceptible to ciprofloxacin (93.3%), gentamicin (46.7%) and trimethoprim-sulfamethoxazole (93.3%) than type III isolates. All MRSA isolates were susceptible to glycopeptides and vancomycin (minimum inhibitory concentrations <2 microg/mL). Pulsed-field gel electrophoresis with SmaI digestion was used to fingerprint these isolates. A total of 9 genotypes with 26 type-subtypes were identified. Genotype A was the most frequent (9 subtypes) indicating that it is epidemic in this hospital. CONCLUSION: After analysis, SCCmec typing could be used to predict drug susceptibility. Specific clones of S. aureus are circulating in hospital and communities in Taiwan.     

Molecular characterization and antibiotic susceptibility of Staphylococcus aureus from a multidisciplinary hospital in Romania

From 2004 to 2005, 60%-72% of invasive Staphylococcus aureus isolates from Romanian hospitals were resistant to methicillin (methicillin-resistant S. aureus [MRSA]), the highest frequency for any European nation. Few reports, however, have addressed the molecular characteristics of S. aureus in Romania. In this study, we utilized spa typing, multilocus sequence typing, staphylococcal cassette chromosome mec (SCCmec) typing, dru typing, pulsed-field gel electrophoresis, and detection of virulence factors to characterize 146 S. aureus strains isolated from 2004 to 2005 at the Clinic County Hospital in Bra?ov. Antibiotic susceptibility patterns for all MRSA isolates and patient demographic data were also obtained. Fifty-six strains (38.4%) were determined to be MRSA by susceptibility testing and SCCmec typing. All MRSA strains were resistant to beta-lactams and tetracycline, but susceptible to nitrofurans, vancomycin, and clindamycin, with inducible clindamycin resistance in 23/28 clindamycin-sensitive/erythromycin-resistant isolates. Molecular typing identified 15 clonal backgrounds (CC 1, 5, 8, 8/239, 9, 15, 20, 22, 25, 30, 45, 80, 97, 101, and 121), only 4 of which were associated with MRSA (CC 1, 8/239, 30, and 80). Spa types 35 (t127, CC 1) and 351 (t030, CC 8/239) accounted for 27.4% and 21.9% of all S. aureus strains, respectively, and 19.6% and 57.1% of all MRSA strains. Both hospital-associated (SCCmec type III) and community-associated (SCCmec type IV) elements were identified within MRSA strains, whereas Panton-Valentine leukocidin was detected in 10 MRSA and 12 methicillin-sensitive S. aureus strains. These results demonstrate the presence of various endemic S. aureus clones within the Clinic County Hospital in Bra?ov, suggestive of ongoing nosocomial and community transmission.     

mecA gene is widely disseminated in Staphylococcus aureus population

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most important causes of hospital infections worldwide. High-level resistance to methicillin is caused by the mecA gene, which encodes an alternative penicillin-binding protein, PBP 2a. To determine the clonal relationships between methicillin-susceptible S. aureus (MSSA) and MRSA, we typed 1,069 S. aureus isolates (493 MSSA isolates and 576 MRSA isolates), collected mainly in North American and European hospitals between the 1960s and the year 2000, using pulsed-field gel electrophoresis and ribotyping. Of 10 widespread S. aureus lineages recognized, 8 had corresponding mecA-positive strains. Multiresistant MRSA strains are found in hospitals worldwide, while unrelated and more susceptible strains represent less than 1% of the MRSA population. This supports the hypothesis that horizontal transfer plays an important role in the dissemination of the mecA gene in the S. aureus population.     

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.     

A high frequency of macrolide-lincosamide-streptogramin resistance determinants in Staphylococcus aureus isolated in South Korea

Genes conferring resistance to one of the macrolide-lincosamide-streptogramin (MLS) antibiotics may confer cross-resistance to others, because they have similar effects on bacterial protein synthesis. In Korea, over 70% of Staphylococcus aureus isolates are methicillin-resistant and erythromycin-resistant methicillin-resistant S. aureus (MRSA) is also prevalent. We investigated the frequency of MLS resistance in erythromycin-resistant S. aureus isolates. A total of 682 isolates of S. aureus were collected in a nationwide antibiotic resistance survey. Susceptibility to erythromycin, clindamycin, and quinupristin/dalfopristin was tested by disk diffusion. In all, 37% of the methicillin-susceptible S. aureus (MSSA) and 97% of the MRSA isolates were resistant to at least one of the MLS antibiotics, whereas all were susceptible to quinupristin/dalfopristin. Out of 518 strains that were resistant to erythromycin, 60 clindamycin-susceptible (30 MSSA, 30 MRSA) and 44 clindamycin-resistant isolates (14 MSSA, 30 MRSA) were selected at random from these strains. Thirteen genes related to MLS resistance were detected in these isolates by PCR. Of the 104 MSSA and MRSA strains tested, 98 harbored one or more erm gene. The most common was erm(A), with erm(C) next. But, msr(A), lnu(A), and mef(A) were rare and no resistance to streptogramin A was encountered.     

Low Prevalence of Community-Acquired Methicillin-Resistant Staphylococcus aureus in Adults at a University Hospital in the Central United States

Community-acquired MRSA (CA-MRSA) is potentially a new emerging pathogen with most strains susceptible to many antimicrobials except for beta-lactam antibiotics. We retrospectively reviewed MRSA isolates during a 20-month study period (January 1998 through August 1999) and investigated those that were clindamycin susceptible. Patients were not considered to harbor CA-MRSA if they had been admitted to a hospital within the preceding 2 years or if their isolate had been obtained more than 72 h after admission. There were 2,817 S. aureus isolates, with 1,071 (38%) being MRSA. Of these 1,071 isolates, 161 were clindamycin susceptible; these were recovered from 81 patients. Of these 81 patients, 20 appeared to have community-acquired strains, but only 2 could be confirmed as having CA-MRSA.     

Characterization of Staphylococcus aureus nasal carriage from children attending an outpatient clinic in Seoul, Korea

Nasal swabs were collected to isolate S. aureus in 296 children, who visited the pediatrics department with a variety of symptoms. Staphylococcus aureus was isolated from 95 children (32.1%). Of the isolates, 18 were methicillin-resistant S. aureus (MRSA) (18.9%). Antimicrobial susceptibility testing was performed for all S. aureus cultured and the molecular characteristics were investigated. Forty-nine spa types were identified among the S. aureus isolates, and were classified into 13 spa groups (A-L). The most prevalent clone (34 isolates, 35.8%) belonged to the spa group B (spa repeat motif, WG/FKAOMQ), which corresponded to sequence type 30 (ST30) and its variants. Sixteen different spa types, within the spa group B, suggested that this group has evolved over a long period of time. In addition, all S. aureus isolates belonging to the spa group B were methicillin-susceptible, indicating that this group might represent successful adaptation of this clone in the community setting with low antibiotic pressure. The most frequently found clone in the MRSA group was spa group C (spa repeat motif, DMGGM) and SCCmec type IVA, which represented half of the MRSA isolates and corresponded to ST72. ST5-MRSA-II, the most prevalent MRSA clone in Korean hospitals, was found in only two isolates. These findings suggest that strains of S. aureus nasal carriage in Korean children visiting an outpatient pediatric department were different from the strains identified in hospital infections.     

Comparison of trends of resistance rates over 3 years calculated from results for all isolates and for the first isolate of a given species from a patient

We compared trends of annual resistance rates calculated from results for all isolates and for the first isolate of Staphylococcus aureus, Klebsiella pneumoniae, and Acinetobacter baumannii per patient over a 3-year period from 2001 through 2003. Antimicrobial susceptibility results of inpatients were extracted from a computerized database. Annual resistance rates of a species were calculated by two methods: (i) from results for all isolates, even those from patients with multiple isolates in a given year and (ii) from results for the first isolate from a patient in a given year, regardless of susceptibility profile or specimen type. Rates of methicillin-resistant S. aureus (MRSA) did not differ among all isolates (79.9, 78.8 and 79.6%; P = 0.86), but decreased for the first isolate per patient (70.2, 65.7, and 64.1%; P = 0.006) over time. Annual duplication rates of methicillin-susceptible S. aureus (MSSA) decreased (39.6, 37.6, and 31.7%; P = 0.01), but those of MRSA increased significantly (64.3, 67.8, and 68.9%; P = 0.004). Rates of cefotaxime-resistant K. pneumoniae did not differ over time by either method, and rates of imipenem-resistant A. baumannii decreased over time by both methods. Duplication rates did not differ for either susceptible or resistant isolates of K. pneumoniae and A. baumannii. The trends in MRSA rate differed by the two methods because of the different proportion of duplicate isolates per year. MRSA rates might be increasingly overestimated for all isolates. These results suggest that the method of calculating results for the first isolate per patient may remove the effect of duplication, allowing the simple and unambiguous analysis of cumulative susceptibility rates.     

In vitro activities of quinupristin/dalfopristin and eight other antimicrobial agents against 360 clinical isolates from Korea

The emergence of multi-drug resistant gram-positive cocci such as methicillin-resistant (MR) staphylococci, vancomycin-resistant (VR) enterococci, and vancomycin-intermediate resistant S. aureus (VISA) has given new urgency to the development of new antimicrobial agents. One of these is quinupristin/dalfopristin (Q/D). We decided to determine the susceptibility of gram-positive cocci isolated at two university hospitals in Seoul to Q/D and compare the results with eight other antimicrobial agents. We investigated 120 isolates of S. aureus including 49 MRSAs and one VISA, 120 isolates of coagulase negative staphylococci (CNS), 64 E. faecalis and 56 E. faecium, including seven strains of VR E. faecium. Minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBCs) for several antimicrobials, including vancomycin and Q/D, were determined by broth microdilution. All S. aureus including VISA were susceptible to Q/D. Q/D MIC90 for both methicillin-susceptible S. aureus (MSSA) and MRSA was 0.25 g/mL. 49 (87.5%) of 56 E. faecium including six of seven VR E. faecium were susceptible to Q/D. E. faecalis were not susceptible to Q/D (only 1.5% susceptible), but were inhibited by ampicillin (94% susceptible) or vancomycin (95%). CNS was susceptible to Q/D (96% susceptible) and vancomycin (100% susceptible). One of 38 staphylococci and two of 17 E. faecium were tolerant to Q/D. In conclusion, Q/D showed excellent activity against all species of gram-positive cocci including MRSA, VISA, and VR E. faecium except E. faecalis, and may provide a valuable option for the treatment of infections caused by these emerging nosocomial pathogens of gram-positive cocci.     

Prevalence and characterization of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus, isolated from bulk tank milk from Minnesota dairy farms

Staphylococcus aureus is a common causative agent of bovine mastitis in dairy herds. The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in hospitals as well as the community is a significant and costly public health concern. S. aureus-related bovine mastitis is a common reason for therapeutic and/or prophylactic use of antibiotics on dairy farms. In this study, herd prevalence of S. aureus, including MRSA, was estimated from bulk tank milk (BTM) from Minnesota farms. A total of 150 pooled BTM samples from 50 farms, collected over 3 seasons (spring, summer, and fall of 2009), were assessed. Herd prevalence of methicillin-susceptible S. aureus (MSSA) was 84%, while MRSA herd prevalence was 4%. A total of 93 MSSA isolates and 2 MRSA isolates were recovered from 150 BTM samples. Antibiotic susceptibility testing of S. aureus isolates showed pansusceptibility in 54 isolates, resistance to a single antibiotic class in 21 isolates, resistance to two antibiotic classes in 13 isolates, and resistance to ≥3 antibiotics classes and thus multidrug resistance in 5 isolates. The two MRSA isolates displayed resistance to β-lactams, cephalosporins, and lincosamides and were multiresistant. Staphylococcal protein A gene (spa) typing identified spa types t529 and t034 most frequently among methicillin-susceptible isolates, while t121 was observed in MRSA isolates. Seven isolates, including the two MRSA isolates, produced staphylococcal enterotoxins B, C, D, and E on overnight culture. MRSA isolates were further genotyped using multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Of the 2 MRSA isolates, one had a composite genotype profile of MLST ST 5-PFGE USA100-unknown spa type, which has been reported among hospital-associated MRSA isolates, while the second isolate carried the MLST ST 8-PFGE USA300-spa type t121 genotype, commonly identified among community-associated MRSA isolates. These results suggest that MRSA genotypes associated with hospitals and community can be isolated from milk at very low rates.     

Staphylococcus aureus susceptibility to innate antimicrobial peptides,beta-defensins and CAP18, expressed by human keratinocytes

The antimicrobial peptides human beta-defensin-1 (hBD1), hBD2, hBD3, and CAP18 expressed by keratinocytes have been implicated in mediation of the innate defense against bacterial infection. To gain insight into Staphylococcus aureus infection, the susceptibility of S. aureus, including methicillin-resistant S. aureus (MRSA), to these antimicrobial peptides was examined. Based on quantitative PCR, expression of hBD2 mRNA by human keratinocytes was significantly induced by contact with S. aureus, and expression of hBD3 and CAP18 mRNA was slightly induced, while hBD1 mRNA was constitutively expressed irrespective of the presence of S. aureus. Ten clinical S. aureus isolates, including five MRSA isolates, induced various levels of expression of hBD2, hBD3, and CAP18 mRNA by human kertinocytes. The activities of hBD3 and CAP18 against S. aureus were found to be greater than those of hBD1 and hBD2. A total of 44 S. aureus clinical isolates, including 22 MRSA strains, were tested for susceptibility to hBD3 and CAP18. Twelve (55%) and 13 (59%) of the MRSA strains exhibited more than 20% survival in the presence of hBD3 (1 microg/ml) and CAP18 (0.5 microg/ml), respectively. However, only three (13%) and two (9%) of the methicillin-sensitive S. aureus isolates exhibited more than 20% survival with hBD3 and CAP18, respectively, suggesting that MRSA is more resistant to these peptides. A synergistic antimicrobial effect between suboptimal doses of methicillin and either hBD3 or CAP18 was observed with 10 MRSA strains. Furthermore, of several genes associated with methicillin resistance, inactivation of the fmtC gene in MRSA strain COL increased susceptibility to the antimicrobial effect mediated by hBD3 or CAP18.     

Prevalence of methicillin-resistant Staphylococcus aureus in eight African hospitals and Malta

Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious therapeutic problem worldwide, and its frequency in most African countries has not been reported. This study was aimed at determining the prevalence and antibiotic susceptibility patterns of MRSA in eight large hospitals (>500 beds) in Africa and Malta, from 1996 to 1997. Susceptibility to methicillin (oxacillin) and to other drugs was determined by E test (AB Biodisk, Solna, Sweden) on a total of 1440 clinical isolates of S. aureus . Methicillin resistance was detected in 213 (15%) of the 1440 isolates tested. The rate of MRSA was relatively high in Nigeria, Kenya, and Cameroon (21–30%), and below 10% in Tunisia, Malta, and Algeria. All MRSA isolates were sensitive to vancomycin, with MICs ≤ 4 mg/L. The isolates were also highly sensitive to ciprofloxacin, except in Kenya, Morocco, and Tunisia, where relative resistance to this drug was noted. Susceptibility to rifampin and fusidic acid seems to be correlated with the clinical use of these compounds. Only 46% of 59 MRSA strains analyzed were susceptible to rifampin, fusidic acid, and ciprofloxacin. The majority (> 60%) of MRSA strains were multiresistant. There is a need to maintain surveillance and control of MRSA infections in Africa.     

Staphylococcal cassette chromosome mec (SCCmec) analysis and antimicrobial susceptibility patterns of methicillin-resistant Staphylococcus aureus (MRSA) isolates in Tehran, Iran

Oxacillin resistance was present in 99 of 277 (36%) consecutive Staphylococcus aureus isolates collected from hospital patients in Tehran during a 15-month period (January 2004-March 2005). The majority of isolates (77/99 = 78%) had been cultured from wounds or blood. The staphylococcal cassette chromosome mec (SCCmec) types and antimicrobial susceptibility patterns of 99 methicillin-resistant S. aureus (MRSA) strains were determined. Disk diffusion and agar dilution methods were used to determine the susceptibility of isolates to antimicrobial agents as instructed by Clinical and Laboratory Standards Institute. The presence of mecA and SCCmec types was determined by PCR and multiplex PCR. All MRSA isolates were susceptible to vancomycin (MIC90 相似文献   

Increased vancomycin MICs for Staphylococcus aureus clinical isolates from a university hospital during a 5-year period

Staphylococcus aureus is one of the most commonly isolated organisms in nosocomial infections. While the prevalence of methicillin-resistant S. aureus (MRSA) continues to increase worldwide, there is concern about an increase in vancomycin MICs among S. aureus strains. The prevalence of MRSA and vancomycin MIC trends in S. aureus from patients in a university hospital were analyzed. Clinical Laboratory Standards Institute (CLSI, formerly NCCLS) reference broth microdilution MIC testing was performed on all clinically relevant S. aureus isolates from January 2000 through December 2004. A total of 6,003 S. aureus isolates were analyzed. No vancomycin-resistant S. aureus isolates were detected. One MRSA isolate had a vancomycin MIC of 8 mug/ml and was confirmed as vancomycin-intermediate S. aureus. Among the 6,002 remaining isolates, a shift in vancomycin MICs from 相似文献   

Methicillin-Resistant Staphylococcus aureus: Comparison of Susceptibility Testing Methods and Analysis of mecA-Positive Susceptible Strains

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for an increasing number of serious nosocomial and community-acquired infections. Phenotypic heterogeneous drug resistance (heteroresistance) to antistaphylococcal beta-lactams affects the results of susceptibility testing. The present study compared the MRSA-Screen latex agglutination test (Denka Seiken Co., Ltd., Tokyo, Japan) for detection of PBP 2a with agar dilution, the VITEK-1 and VITEK-2 systems (bioMérieux, St. Louis, Mo.), and the oxacillin agar screen test for detection of MRSA, with PCR for the mecA gene used as the "gold standard" assay. Analysis of 107 methicillin-susceptible S. aureus (MSSA) isolates and 203 MRSA isolates revealed that the MRSA-Screen latex agglutination test is superior to any single phenotype-based susceptibility testing method, with a sensitivity of 100% and a specificity of 99.1%. Only one isolate that lacked mecA was weakly positive by the MRSA-Screen latex agglutination test. This isolate was phenotypically susceptible to oxacillin and did not contain the mecA gene by Southern blot hybridization. The oxacillin agar screen test, the VITEK-1 system, the VITEK-2 system, and agar dilution showed sensitivities of 99.0, 99.0, 99.5, and 99%, respectively, and specificities of 98.1, 100, 97.2, and 100%, respectively. The differences in sensitivity or specificity were not statistically significant. Oxacillin bactericidal assays showed that mecA- and PBP 2a-positive S. aureus isolates that are susceptible to antistaphylococcal beta-lactams by conventional methods are functionally resistant to oxacillin. We conclude that the accuracy of the MRSA-Screen latex agglutination method for detection of PBP 2a approaches the accuracy of PCR and is more accurate than any susceptibility testing method used alone for the detection of MRSA.     

Inducible clindamycin resistance and nasal carriage rates of Staphylococcus aureus among healthcare workers and community members

BackgroundNasal carriage of Staphylococcus aureus is becoming an increasing problem among healthcare workers and community individualsObjectivesTo determine the prevalence of methicillin-resistant S. aureus (MRSA) nasal colonization and inducible clindamycin resistance (ICR) of S. aureus among healthcare workers at Soba University Hospital and community members in Khartoum State, Sudan.MethodsFive hundred nasal swabs samples were collected during March 2009 to April 2010. Isolates were identified using conventional laboratory assays and MRSA determined by the disk diffusion method. The D-test was performed for detection of ICR isolates with Clinical Laboratory Standard Institute guidelines.ResultsOf the 114 S. aureus isolated, 20.2% represented MRSA. The occurrence of MRSA was significantly higher among healthcare worker than community individuals [32.7% (18/55) vs. 6.9% (5/59)] (p=0.001). Overall the 114 S. aureus isolates tested for ICR by D-test, 29 (25.4%) yielded inducible resistance. Significantly higher (p=0.026) ICR was detected among MRSA (43.5%) than methicillin-susceptible S. aureus (MSSA) (20.9%).ConclusionMRSA nasal carriage among healthcare workers needs infection control practice in hospitals to prevent transmission of MRSA. The occurrence of ICR in S. aureus is of a great concern, D- test should be carried out routinely in our hospitals to avoid therapeutic failure.