Oxacillin Disk Diffusion Research Articles

Evaluation of Susceptibility Test Breakpoints Used to Predict mecA-Mediated Resistance in Staphylococcus Pseudintevmedius Isolated from Dogs

Clinical and Laboratory Standards Institute interpretive breakpoints for in vitro susceptibility tests that predict mecA-mediated oxacillin resistance in Staphylococcus pseudintermedius isolates from animals have been changed twice in the past decade. Moreover, there are no counterpart recommendations for human isolates of S. pseudintermedius. Individual medical and veterinary laboratories variably use interpretive breakpoints identical to those recommended for use with Staphylococcus aureus or identical to those recommended for use with coagulase-negative staphylococci. The purpose of the current study was to examine correlations between oxacillin disk diffusion, oxacillin gradient diffusion, oxacillin microbroth dilution, and cefoxitin disk diffusion tests used to predict mecA-mediated resistance in S. pseudintermedius and to retrospectively estimate, from disk diffusion zone diameter measurements, the prevalence and rate of increase of oxacillin resistance among canine S. pseudintermedius isolates submitted to a veterinary teaching hospital laboratory. Oxacillin disk diffusion zone diameters of <or=17 mm and oxacillin minimum inhibitory concentrations of >or=0.5 microg/ml were highly correlated with detection of mecA in canine S. pseudintermedius isolates by polymerase chain reaction. MecA-mediated resistance among S. pseudintermedius isolates from dogs increased from less than 5% in 2001 to near 30% in 2007. More than 90% of the methicillin-resistant S. pseudintermedius isolates in 2006 and 2007 were also resistant to representatives of >or=4 additional antimicrobial drug classes. Cefoxitin disk diffusion with the resistance breakpoint set at <or=24 mm significantly underestimated the presence of mecA in S. pseudintermedius.

Antimicrobial activity of tigecycline and comparative agents against clinical isolates of staphylococci and enterococci from ICUs and general hospital wards at three Swedish university hospitals

The activities of tigecycline and comparative agents on staphylococci and enterococci isolated from patients at general hospital wards (GHWs) and intensive care units (ICUs) at 3 university hospitals in Sweden were investigated. Oxacillin disc diffusion and minimal inhibitory concentration with E-test were used. The presence of mecA, vanA or vanB genes was determined with PCR. Statistically significant higher incidence of clindamycin, fusidic acid, rifampicin and multidrug-resistant CoNS was found at ICUs compared to GHWs. Resistance rates were low among S. aureus. Tigecycline, linezolid and vancomycin were the only agents with high activity against methicillin-resistant S. aureus and multidrug-resistant CoNS. Resistance rates were low among E. faecalis, except for high-level gentamicin-resistant (HLGR) E. faecalis. E. faecium showed high resistance rates to ampicillin, piperacillin/tazobactam and imipenem. The HLGR rates among E. faecium were lower than the rates for E. faecalis. Tigecycline and linezolid were the only drugs with high activity against all enterococci including vancomycin-resistant enterococci. No statistically significant differences in susceptibility rates were found between the ward levels for S. aureus and enterococcal isolates and no statistically significant differences were found between the hospitals.

COMPARISON OF CEFOXITIN DISC DIFFUSION TEST, OXACILLIN SCREEN AGAR, AND PCR FOR mecA GENE FOR DETECTION OF MRSA

Cefoxitin is a potent inducer of the mecA regulatory system. It is being recommended for detection of methicillin resistance in Staphylococcus aureus (MRSA) when using disk diffusion testing. The aim of our study was to evaluate the efficacy of cefoxitin disc diffusion test to characterize MRSA and compare it with oxacillin agar screening and detection of mecA gene by PCR. Materials and Methods: Fifty strains of S. aureus isolated from clinical samples were used in the study. Routine antibiotic susceptibility testing was performed including oxacillin disk. Oxacillin screen agar plates with 4% NaCl and 6 µg/ml of oxacillin were inoculated and interpreted as per standard guidelines. Cefoxitin disc diffusion test was performed using 30 µg disc and zone sizes were measured. PCR for amplification of the mecA gene was performed. Results: Out of the 50 isolates, 28 were found to be methicillin resistant by oxacillin disc diffusion test, 30 were resistant by oxacillin screen agar method, and 32 were resistant with cefoxitin disc diffusion. For these 32 isolates mecA gene was positive. Conclusion: Results of cefoxitin disc diffusion test is in concordance with the PCR for mecA gene. Thus, the test can be an alternative to PCR for detection of MRSA in resource constraint settings.

Cefoxitin disc diffusion test for detection of meticillin-resistant staphylococci

Staphylococci are the main causative agents of nosocomial diseases. Over the last few years, the increase in the number of meticillin-resistant (MR) staphylococci has become a major clinical problem. Accuracy and promptness in the detection of meticillin resistance are of key importance in ensuring the correct antibiotic treatment in infected patients and control of MR staphylococci in the hospital environment. This study evaluated the accuracy of a cefoxitin disc diffusion (DD) test for the detection of meticillin resistance in staphylococci. A total of 144 clinical isolates [97 Staphylococcus aureus and 47 coagulase-negative staphylococci (CoNS)] were tested using a mecA gene PCR, a DD test (oxacillin, 1 mug disc; cefoxitin, 30 mug disc), determination of oxacillin MIC by agar dilution (AD), and an oxacillin screen agar test at oxacillin concentrations of 4 and 6 mug ml(-1). Of the 97 S. aureus and 47 CoNS isolates, 73 (75.26 %) and 30 (63.83 %), respectively, were mecA-positive. The sensitivity and specificity of the cefoxitin DD test were 94.44 and 95.83 %, respectively, for S. aureus and 80 and 100 %, respectively, for CoNS. The oxacillin DD method was 100 % sensitive and 58.33 % specific for S. aureus, and 86.67 % sensitive and 70.59 % specific for CoNS. The AD test was highly sensitive (98.63 %) and specific (98.53 %) for S. aureus and CoNS (83.33 % sensitive and 94.12 % specific). The cefoxitin DD test for meticillin-resistance detection was more specific but less sensitive than the oxacillin DD test. Use of DD tests for both cefoxitin and oxacillin can help in more accurate prediction of meticillin resistance. Centres that are not equipped to carry out PCR can use AD methods for confirmation of meticillin resistance, especially in oxacillin-resistant and cefoxitin-sensitive cases.

Prevalence of Staphylococcus aureus Nasal Colonization in Emergency Department Personnel

Nasal colonization with Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) can precede infection in patients and contacts. Although general population S aureus/MRSA rates are well described, the prevalence of S aureus and MRSA nasal colonization in emergency department health care workers is not defined. We seek to determine the prevalence of S aureus and MRSA nasal colonization among ED health care workers without evidence of an active site of staphylococcal infection and identify variables associated with colonization. We prospectively studied a convenience sample of ED health care workers from 5 urban teaching hospitals in Pittsburgh, PA. Each participant completed a questionnaire and nasal culturing. We tested susceptibility with the oxacillin disc diffusion method. We analyzed data with descriptive statistics and univariate regression, with alpha set at 0.05. Of 255 subjects, 23% were physicians; 62% were nurses, nursing assistants, or patient care technicians; and 15% were clerical staff or social service workers. Of 81 (31.8%) S aureus isolates, 11 (13.6%) were MRSA, an overall prevalence of 4.3%. All positive MRSA samples were from nurses, nursing assistants, or patient care technicians. No other covariate had an association with S aureus or MRSA colonization. In this urban ED health care worker population, the prevalence of S aureus is similar but MRSA nasal colonization is higher than previously reported estimates in the general population of the United States. Physicians and nonpatient contact ED health care workers did not have MRSA colonization.

Comparison of polymerase chain reaction and conventional methods in detecting methicillin-resistant Staphylococcus aureus

Accurate and rapid detection of methicillin-resistant Staphylococcus aureus is very important in a clinical laboratory setting to avoid treatment failure. Conventional methods were compared against the gold standard polymerase chain reaction (PCR) technique to determine the best combination of the routine procedures. Methicillin resistance was investigated in 416 clinical Staphylococcus aureus isolates by PCR, oxacillin agar screening (OAS), oxacillin disk diffusion (ODD) and cefoxitin disk diffusion (CDD) methods. Two hundred and ten (51%) out of 416 S. aureus strains were found to be mecA-positive by PCR. Sensitivity and specificity of the ODD, CDD and OAS methods were detected as follows: 100% and 89%, 99.50% and 100%, and 99.50% and 100%, respectively. Combining the ODD and CDD methods could be a good choice for detecting methicillin resistance in S. aureus strains where mecA PCR cannot be performed.

Identification of Methicillin Resistant Staphylococcus aureus (MRSA) and Methicillin Resistant Coagulase-Negative Staphylococcus (CoNS) in Clinical Settings

To evaluate different methods for the identification of methicillin-resistant Staphylococci and their reliability, 112 Staphylococcal isolates (32 Staphylococcus aureus isolates and 80 coagulase negative Staphylococci isolates “CoNS”) were collected from 118 nasal swab cultures and were subjected to three methods to detect oxacillin susceptibility of the isolates. The three methods were oxacillin disk diffusion method; the Epsilometer- test (E-test) and polymerase chain reaction (PCR). For the S. aureus strains, the E-test and the PCR methods showed discrepant results in two isolates (6.25%); that showed susceptible patterns with the E-test, but were resistant by the disk diffusion method. Both isolates were negative for the presence of mecA gene. Seven (8.75%) out of the 80 CoNS isolates showed conflicting results where four isolates showed resistance with the disk diffusion and the E-test methods, and had negative mecA gene by PCR. Three of the 7 CoNS with the conflicting results showed a susceptible pattern to oxacillin by the E-test method, while the PCR method showed the presence of mecA gene. We concluded that combination of molecular and conventional methods should be used to assess methicillin resistance of Staphylococci in clinical practices.

Evaluation of phenotypic and molecular methods for screening and detection of methicillin-resistant Staphylococcus aureus

An in-house PCR was compared with the GenoType MRSA and the MRSA EVIGENE tests, both of which corresponded 100% with the results of in-house PCR. The cefoxitin disk diffusion method was superior to both the oxacillin disk diffusion and minimum inhibitory concentration tests for predicting the mecA status.

Evalution of methods for the detection of MRSA in patients of Botucatu Medical School Hospital, São Paulo, Brazil

Oxacillin is the main drug of choice for the treatment of S. aureus infections. However, S. aureus resistance to oxacillin has become a major problem in the recent decades. The study aimed assess the rates of oxacillin resistance in S. aureus samples obtained at the Botucatu Medical School Hospital, UNESP, and to compare phenotypic techniques for the detection of MRSA against the gold standard method (mecA gene detection) in these samples. A total of 102 samples, previously isolated between 2002 and 2006, and kept at the Culture Collection of the Department of Microbiology and Immunology, in the Botucatu Biosciences Institute, UNESP, were included. Oxacillin resistance was assessed by oxacillin and cefoxitin disk diffusion and agar dilution tests, screening tests using Mueller-Hinton agar with 6 μg/mL of oxacillin and 4% NaCl, E-test, and mecA gene detection. Of the samples analyzed, 46 (45.1%) were mecA-positive. Oxacillin disk sensitivity and specificity were 86.9% and 91.1%, respectively. Cefoxitin disk sensitivity and specificity were respectively 91.3% and 91.1%. The screening test with the cefoxitin disk showed almost the same level of sensitivity (91.3%) and specificity (91.1%). With E-test strips, sensitivity was higher (97.8%) and specificity was comparable to that found with the other methods (91.1%). Ninety-three percent of the samples produced β- lactamase and five of them were mecA-negative. There was a gradual increase in the number of oxacillin-resistant S. aureus samples between 2002 and 2004. However, from 2004 to 2006, the number of resistant samples dropped from 55% of MRSA in 2004, to 45% in 2005 and 34.6% in 2006. The data obtained reveal that, among phenotypic methods, the E-test yielded the best results, with higher sensitivity levels when compared to the other methods. The decreased resistance rate observed over the most recent years may be explained by the rational use of antimicrobial agents associated with good practices in the control of hospital infection, or may be related to the diminished use of oxacillin as a treatment option.

Comparison of mecA Polymerase Chain Reaction With Phenotypic Methods for the Detection of Methicillin-Resistant Staphylococcus aureus

In the present study, several conventional methods to detect methicillin-resistant Staphylococcus aureus (MRSA) were compared with polymerase chain reaction (PCR) detection of mecA gene-positive isolates. Cefoxitin E-test was also evaluated as a possible phenotypic method of MRSA detection. Oxacillin agar screen and PBP2' latex agglutination methods were found to be more sensitive than oxacillin and cefoxitin disk-diffusion methods. Cefoxitin disk diffusion was found to be the most specific. A combination of oxacillin agar screening with cefoxitin disk diffusion, or oxacillin disk diffusion with PBP2', improved sensitivity and specificity. Cefoxitin E-test with the current break points had low sensitivity and specificity (33.3% and 75%, respectively) for the detection of MRSA. However, changing the break points to <or= 4 microg/ml and to >or= 6 microg/ml for sensitive and resistant, respectively, greatly improved both. Changing the 30-microg cefoxitin disk-diffusion break points to <or= 21 mm for resistant slightly improved sensitivity but had no effect on specificity. It was therefore concluded that the use of more than one screening method is necessary to detect all MRSA isolates in clinical settings.

ANTIMICROBIAL RESISTANCE IN INVASIVE AND COLONISING STREPTOCOCCUS PNEUMONIAE IN NORTH INDIA

The present study was done to detect the antibiotic resistance in S. pneumoniae. One hundred twenty S. pneumoniae isolates from clinical specimens and 50 from nasopharyngeal sites were subjected to antimicrobial susceptibility testing by Kirby Bauer disk diffusion method and minimum inhibitory concentration (MIC) determination for penicillin and cefotaxime non-susceptible isolates. A total of 22 isolates (18.3%) from clinical sites and eight (16%) from nasopharyngeal sites showed decreased susceptibility to penicillin by oxacillin disk diffusion test. MICs of 26 of these resistant strains ranged from 0.12-1 µg/mL (intermediate resistance) by broth dilution and E test. Only four isolates, two from sputum and two from nasopharyngeal swabs, showed MIC of 2 µg/mL (complete resistance). However, MIC of two cefotaxime resistant isolates (by disk diffusion) was in the susceptible range (0.5 µg/mL). Highest antimicrobial resistance was seen to cotrimoxazole (55.2%) and tetracycline (61.2%). Antimicrobial resistance to cotrimoxazole and tetracycline was much more in clinical isolates than colonizing isolates. Multi-drug resistant phenotype was detected in 76.9% (20 of 26) of isolates that were intermediately sensitive to penicillin and 50% (2 of 4) of penicillin resistant isolates (co-resistant to tetracycline and cotrimoxazole). Routine screening for antibiotic susceptibility is recommended for clinical isolates of pneumococci. Strains with reduced susceptibility to penicillin should be subjected to MIC determination to detect relative resistance or true resistance as such strains are associated with increased virulence.The choice of antibiotics should be guided by the prevalence of local resistance patterns of pneumococci.

The cefoxitin disk test – what a clinical microbiologist needs to know

Accurate prediction of mecA- mediated resistance in Staphylococcus spp. has been a challenge for the clinical microbiologist ever since the discovery of methicillin resistance. In 2004, a cefoxitin disk diffusion test was recommended by the Clinical and Laboratory Standards Institute (formerly NCCLS) for prediction of mecA- mediated resistance in both Staphylococcus aureus and coagulase-negative staphylococci because of its high sensitivity and specificity compared to PCR tests for mecA. It is also much easier to read that the oxacillin disk diffusion test, not requiring close scrutiny of the zone of inhibition or 24 h of incubation. The test is used as a surrogate for oxacillin testing. In other words, when cefoxitin is tested, the results are applied to oxacillin and oxacillin is reported. For resistant strains, the results are applied not only to oxacillin, but also to all other β-lactam agents, which must also be reported as resistant if they are reported. There has been some confusion among clinical microbiologists about when and how the test should be used. We answer some of the questions that have been raised about the test.

Detection of mecA-mediated resistance using reference and commercial testing methods in a collection of Staphylococcus aureus expressing borderline oxacillin MICs

Phenotypic methods for detecting mecA-mediated resistance in Staphylococcus aureus include both oxacillin and cefoxitin susceptibility tests; many laboratories perform multiple tests. Conflicting oxacillin and cefoxitin susceptibility results are most likely to occur for isolates that either have reduced susceptibility to oxacillin by a non– mecA-mediated mechanism or are mecA positive but are very heteroresistant. To understand the performance of oxacillin and cefoxitin tests for such isolates, we tested 135 S. aureus isolates using either cefoxitin or oxacillin and compared the results with mecA polymerase chain reaction. These strains either expressed borderline oxacillin MICs (1–4 μg/mL) and had undetermined mecA status or were mecA positive but were not detected by oxacillin broth microdilution (BMD) or disk diffusion (DD) in original testing. For 24-h readings, performance of cefoxitin tests (sensitivity/specificity) were DD (99/100), Etest using ≤6 μg/mL as susceptible (99/98), and Phoenix MIC using ≤4 μg/mL as susceptible (98/100). Using 6 μg/mL of cefoxitin as a screen test in both BMD and agar dilution also worked well (98/98—100). Sensitivity/specificity of oxacillin methods were oxacillin agar screen (BBL: 80/86; Remel, Lenexa, KS: 85/50), DD (91/59), BMD (85/88), MicroScan (89/96), VITEK Legacy (82/93), VITEK 2 (91/73), and Phoenix, (67/96). These results suggest that a cefoxitin test can be used alone to predict mecA-mediated resistance in S. aureus.

Staphylococcus aureus colonization among nursing home residents in a large Finnish nursing home

We studied colonization with methicillin-resistant and -sensitive Staphylococcus aureus (MRSA, MSSA) in the second largest nursing home in Finland, in which the residents volunteered had their nostrils, throats, perineums, skin lesions, and catheter exit sites swabbed, and catheter urines cultured. The specimens were cultured onto non-selective and selective agar, with or without enrichment in salt-containing trypticase soy broth (TSB). S. aureus was identified by routine methods, methicillin resistance was detected by oxacillin and cefoxitin disk diffusion and MIC E-tests, and GenoType MRSA -test was used for mecA gene confirmation. A total of 663 cultures were obtained from 213 residents. Of those, 165 specimens (25%) from 94 residents (44%) were positive for S. aureus, and 3 specimens (0.4%) from 2 (0.9%) residents were positive for MRSA. Of the 165 S. aureus isolates, 31 (19%) from 25 (27%) residents were found only from sites other than nostrils (30 MSSA and 1 MRSA). TSB enrichment detected additional 33 (5%) S. aureus isolates (32 MSSA and 1 MRSA), resulting in 8 (5%) additional residents. None of the MRSA strains would have been found if only nostrils and throat had been screened, and no enrichment broth had been used.

Antimicrobial resistance in invasive and colonising &lt;i&gt;Streptococcus pneumoniae&lt;/i&gt; in North India

The present study was done to detect the antibiotic resistance in S. pneumoniae. One hundred twenty S. pneumoniae isolates from clinical specimens and 50 from nasopharyngeal sites were subjected to antimicrobial susceptibility testing by Kirby Bauer disk diffusion method and minimum inhibitory concentration (MIC) determination for penicillin and cefotaxime non-susceptible isolates. A total of 22 isolates (18.3%) from clinical sites and eight (16%) from nasopharyngeal sites showed decreased susceptibility to penicillin by oxacillin disk diffusion test. MICs of 26 of these resistant strains ranged from 0.12-1 microg/mL (intermediate resistance) by broth dilution and E test. Only four isolates, two from sputum and two from nasopharyngeal swabs, showed MIC of 2 microg/mL (complete resistance). However, MIC of two cefotaxime resistant isolates (by disk diffusion) was in the susceptible range (0.5 microg/mL). Highest antimicrobial resistance was seen to cotrimoxazole (55.2%) and tetracycline (61.2%). Antimicrobial resistance to cotrimoxazole and tetracycline was much more in clinical isolates than colonizing isolates. Multi-drug resistant phenotype was detected in 76.9% (20 of 26) of isolates that were intermediately sensitive to penicillin and 50% (2 of 4) of penicillin resistant isolates (co-resistant to tetracycline and cotrimoxazole). Routine screening for antibiotic susceptibility is recommended for clinical isolates of pneumococci. Strains with reduced susceptibility to penicillin should be subjected to MIC determination to detect relative resistance or true resistance as such strains are associated with increased virulence. The choice of antibiotics should be guided by the prevalence of local resistance patterns of pneumococci.

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 μg/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.

Evaluation of different methods to detect oxacillin resistance in Staphylococcus aureus and their clinical laboratory utility

Methicillin-resistant Staphylococcus aureus strains (MRSA) are major human pathogens responsible for nosocomial and serious community-acquired infections worldwide. Methicillin resistance in S. aureus is primarily mediated by a modified penicillin-binding protein (PBP2a) encoded by the mecA gene, which is not present in susceptible strains [1–3]. Several additional gene products, such as those encoded by fem (factor essential for methicillin resistance) genes and auxiliary genes, can also contribute to MRSA resistance [1, 2, 4]. Rapid and accurate identification of MRSA is necessary for therapeutic and epidemiological purposes. Currently, several methods are available for detecting oxacillin resistance including phenotypic methods, such as disk diffusion, E-test and broth dilution tests, and genotypic methods that detect the mecA gene [1, 5, 6]. PCR amplification of the mecA gene has been used successfully and is presently considered the gold standard for detecting methicillin resistance in S. aureus [1]. Detection of oxacillin resistance using phenotypic methods is problematic because of variation in the degree of mecA gene expression in heterogeneous populations of MRSA; thus, routine oxacillin disk diffusion tests often fail to detect heterogeneous MRSA populations. The Clinical and Laboratory Standards Institute (formerly the National Committee for Clinical Laboratory Standards) has consequently recommended use of the cefoxitin disk diffusion test rather than the oxacillin disk diffusion test [7]. In the study presented here, the performance and utility of several different methods for detecting MRSA was evaluated. PCR for detecting the mecA gene was used as the gold standard assay against which we compared oxacillin and cefoxitin disk diffusion tests and the GenoType MRSA molecular genetic assay (Hain Lifescience, Nehren, Germany) for detecting MRSA. The objective was to determine whether these disk diffusion tests and GenoType MRSA could detect all MRSA as accurately as mecA PCR and to evaluate their suitability as routine methods for detecting oxacillin resistance in S. aureus in clinical microbiology laboratories. GenoType MRSA is a new DNA-based strip assay kit designed to detect oxacillin resistance in S. aureus. Its performance is based on amplification of a 71 bp fragment of the mecA gene and an 85 bp sequence highly specific for S. aureus and subsequent hybridization of the denatured amplicons to their complementary sequences coated on DNA strips. A total of 224 isolates of S. aureus collected from patients of Suleyman Demirel University Hospital were included in this study. The isolates were collected from different clinical specimens and at different times. All strains were identified using biochemical procedures. Reference strains included ATCC33591 (MRSA) and ATCC29213 (MSSA). Disk diffusion tests were performed for each of the 224 isolates using the following method recommended by the National Committee for Clinical Laboratory Standards [7]: 1 μg oxacillin disks (Oxoid, Basingstoke, UK) were incubated at 35°C and at 30°C on Mueller-Hinton agar (Difco, Detroit, MI, USA) with and without 2% NaCl, and 30 μg cefoxitin disks (Oxoid) were incubated at 35°C on Mueller-Hinton agar. After incubation, inhibition zone sizes for oxacillin and cefoxitin were noted and compared. For cefoxitin, isolates showing a zone of inhibition of ≤19 mm were considered Eur J Clin Microbiol Infect Dis (2006) 25:410–412 DOI 10.1007/s10096-006-0153-8

Evaluation of a cefoxitin disk diffusion test for the detection of mecA-positive methicillin-resistant Staphylococcus saprophyticus

In order to validate the current Clinical and Laboratory Standards Institute (CLSI) criteria for the detection of mecA-mediated resistance in Staphylococcus saprophyticus, 101 clinical isolates, including 8 mecA-positive isolates, were investigated. All the isolates were in the range of the resistant category for coagulase-negative staphylococci with the 1 μg oxacillin disk diffusion method and agar dilution method, despite 93 isolates (92%) being mecA-negative. On the other hand, the 30 μg cefoxitin disk diffusion method showed clearly distinguishable zone diameters between the mecA-positive and -negative isolates. However, four of the mecA-negative isolates that would be considered resistant were false positive, and the current interpretive criteria of the CLSI may thus require reconsideration. This study suggests that the cefoxitin disk diffusion method could be more suitable than the oxacillin disk diffusion method for detecting mecA-mediated resistance in S. saprophyticus.

Detection of Methicillin-Resistant Staphylococcus aureus Using mecA/nuc Genes and Antibiotic Susceptibility Profile of Malaysian Clinical Isolates

The aim of this study is to compare methicillin-resistant Staphylococcus aureus (MRSA) detection methods and to generate antibiogram profile of S. aureus clinical isolates from two teaching hospitals in Malaysia including three reference isolates from American Type Culture Collection (ATCC). The mecA/nuc gene PCR amplification, spot inoculation test and oxacillin disc diffusion test were applied to compare its MRSA detection abilities. No disagreement between the three methods was observed. From 29 bacterial isolates (including the ATCC strains) tested, 19 isolates were confirmed as S. aureus with 14 isolates exhibiting multidrug-resistance. All isolates are still susceptible to vancomycin as indicated by the E-test result. Current biochemical tests are comparable with the molecular detection method for MRSA used in this study while multidrug-resistance traits are present in both MRSA and MSSA clinical isolates. Presently, mupirocin seems to be the best alternative for vancomycin against multidrug-resistant S. aureus infections in Malaysia. Susceptibility profile of 19 S. aureus isolates acquired from two teaching hospitals and ATCC towards 16 selected antibiotics was analyzed and an antibiogram was generated. Findings also indicated resistance against many of the available antibiotics and thus an urgent need to search for alternative antibiotics.

Susceptibility of Streptococcus pneumoniae to penicillin in the state of Minas Gerais, Brazil from 1997-2004

We monitored the susceptibility to penicillin of invasive strains of Streptococcus pneumoniae in two reference laboratories; 502 positive cultures from patients with an active invasive infectious process were analyzed. Streptococcus pneumoniae was identified through conventional procedures, and the oxacillin disc diffusion method was used to check for penicillin susceptibility. Statistical analysis included calculations of the frequency distribution, with 95% confidence intervals (CI), as well as chi-square tests and chi-square for linear trend for temporal analysis of susceptibility. The bacterium was isolated from patients less than a year old (40.7% of the isolates), from infants (55.9%), and from individuals less than 15 years old (64.4%). The majority (88.2%, 95% CI = 85.5%-91.1%) of the 502 isolates were susceptible to penicillin. There was no significant temporal trend of elevation of resistance rate during the study period (p=0.56). We conclude that resistance of S. pneumoniae to penicillin is not yet an important clinical-epidemiological concern in the State of Minas Gerais. To provide necessary support for the adoption of therapeutic and prophylactic measures, epidemiological surveillance should be implemented at a national level to monitor the profile of susceptibility/resistance of S. pneumoniae to penicillin and other antimicrobials.