Molecular characterization of resistance to rifampicin in clinical isolates of Neisseria meningitidis

Among 3904 meningococcal isolates collected between October 2002 and June 2007 by the French Meningococcal Reference Centre, eight (0.20%) were resistant to rifampicin (Rif-R; MIC >1 mg/L) and 27 (0.69%) were intermediate-resistant to rifampicin (Rif-I; MICs between 0.38 mg/L and 1 mg/L) according to the E-test method. The MICs determined by agar dilution were lower, eliminating the E-test intermediate category. All Rif-R isolates had mutations in the rpoB gene, resulting in substitutions at or near amino acid position 552, which were absent in non-resistant isolates. These data suggest that a rifampicin clinical breakpoint of 1.0 mg/L should be adopted for Neisseria meningitidis .     

Antibiotic susceptibility among Staphylococcus epidermidis isolated from prosthetic joint infections,with focus on doxycycline

In recent years, coagulase‐negative staphylococci such as Staphylococcus epidermidis have gained importance as nosocomial pathogens, especially in immunocompromised patients and prosthetic joint infections (PJIs). These infections are often long lasting and difficult to treat due to the production of bacterial biofilm and the transformation of the bacteria into a stationary growth phase. Rifampicin is able to penetrate the biofilm, but to reduce the risk of development of rifampicin resistance it should be used in combination with an additional antibiotic. In this study we used Etest to investigate the antimicrobial susceptibility of 134 clinical isolates of S. epidermidis obtained from PJIs to six oral antibiotics: doxycycline, rifampicin, linezolid, fusidic acid, clindamycin, and ciprofloxacin. We also performed synergy testing on doxycycline in combination with each of the remaining antibiotics. Ninety‐three (69%) of the 134 isolates were susceptible to doxycycline, 94/134 (70%) to rifampicin, 56/134 (42%) to clindamycin, 25/134 (19%) to ciprofloxacin, 81/134 (60%) to fusidic acid, and 100% to linezolid. Thirty‐two (80%) of the 40 isolates not fully susceptible to rifampicin were susceptible to doxycycline. Doxycycline in combination with each of the other investigated antibiotics exerted an additive effect on nearly half of the isolates, with the exception of clindamycin, which displayed an even higher percentage of additive effect (69%). To conclude, as the majority of the S. epidermidis isolates were susceptible to doxycycline, this antimicrobial agent may provide a potential alternative for combination therapy together with rifampicin.     

Isolation of methicillin-resistant coagulase-negative staphylococci from chickens.

Methicillin-resistant coagulase-negative staphylococci were isolated from the nares and skin of 1- to 8-week-old healthy chickens in three flocks from a farm. Isolation of methicillin-resistant coagulase-negative staphylococci was positive for 72 (25.7%) of the 280 chickens tested, with the frequency varying from 2.2 to 100% according to flock. A total of 45 appropriate isolates were selected and subjected to identification. Of the 45 methicillin-resistant coagulase-negative staphylococcal isolates selected, 37 were identified as Staphylococcus sciuri, 5 were identified as Staphylococcus epidermidis, and 3 were identified as Staphylococcus saprophyticus. The distribution of the species was different among the flocks. Comparative analysis of the SmaI-digested chromosomal DNA by pulsed-field gel electrophoresis revealed that the isolates could have originated from a single clone of each of S. sciuri and S. saprophyticus and three clones of S. epidermidis. By two methods based on the PCR technique, the mecA gene was detected in all five representative isolates of each methicillin-resistant coagulase-negative staphylococcal clone. The nucleotide sequence of a PCR fragment obtained from an isolate of S. sciuri was completely identical to the corresponding region of mecA genes reported in human methicillin-resistant Staphylococcus aureus isolates and Staphylococcus epidermidis isolates. The representative methicillin-resistant coagulase-negative staphylococcal isolates were resistant to many beta-lactam antibiotics, and some isolates were also resistant to macrolide and aminoglycoside antibiotics. This is the first evidence of the existence of methicillin-resistant coagulase-negative staphylococci from animals possessing the mecA gene.     

Choice of the NaCl concentration for optimizing the detection of methicillin resistance in Staphylococcus using the gel diffusion method

The detection of oxacillin resistance is evaluated by the disk diffusion method in a collection of 374 Staphylococcus sp strains. The disk diffusion assay is performed with 5-microgram oxacillin disk and a 10(8) CFU/mL inoculum on Mueller-Hinton agar plates supplemented with 2 or 5% NaCl and incubated at 37 degrees C for 24 h. Strains are considered resistant in accordance to the French recommendations (any growth around the disk is observed). The detection of mecA gene is performed by PCR almost for resistant and discordant strains. Results are concordant for 246 of 256 Staphylococcus aureus strains (182 susceptible and 64 resistant strains) and for 105 of 118 S. epidermidis isolates tested (37 susceptible and 68 resistant strains). Six mecA-negative strains (3 S. aureus and 3 S. epidermidis) give false resistant results on agar with 2 and 5% NaCl. Seventeen isolates are discordant on 5% NaCl: 7 mecA-negative S. aureus strains are susceptible on 2% NaCl agar but resistant at 5% (3% false-positive results), 10 mecA-positive S. epidermidis strains are resistant on 2% NaCl agar but susceptible at 5% NaCl (5% false-negative results). The detection of meticillin resistance is improved on agar supplemented with 2% NaCl.     

Detection of methicillin resistance in coagulase-negative staphylococci by cefoxitin disc diffusion and oxacillin Etest. A study of consecutive bacteraemia isolates

Coagulase-negative staphylococci (CoNS) are a significant cause of nosocomial bacteraemia and their susceptibility to beta-lactamase-stabile penicillins is unpredictable. To ensure appropriate antibiotic therapy reliable methods for detection of methicillin resistance (MR) are needed. The objectives of this study were to determine the frequency of MR in a set of CoNS from cases of monomicrobial bacteraemia and to evaluate two phenotypic assays for detection of MR, the 10 microg cefoxitin disk test on Iso-Sensitest agar using a semiconfluent inoculum and the oxacillin Etest. MR was determined by a commercial genomic mecA assay. Of 110 CoNS, 75 were mecA positive and 35 mecA negative. Using interpretive zone diameters R < 22 mm and S > or = 27 mm, the cefoxitin disk test had a sensitivity and specificity of 100%. A correct prediction was obtained for 86 isolates, while 23 were indeterminate (> or = 22 mm; <27 mm). Using CLSI's guidelines, sensitivity and specificity of the oxacillin Etest were 100% and 80%, respectively. A correct prediction was obtained for 102 isolates, while 7 mecA negative isolates were classified as resistant. Thus, the cefoxitin disk test and the oxacillin Etest performed with high accuracy and both seem to be suitable for routine use.     

Methicillin-resistant Staphylococcus saprophyticus in Sweden carries various types of staphylococcal cassette chromosome mec (SCCmec)

Staphylococcus saprophyticus is a common cause of uncomplicated urinary tract infections and is usually susceptible to the antimicrobial agents used for their treatment. However, S. saprophyticus resistant to β-lactam antibiotics and carrying mecA has been reported. Eight Swedish isolates of mecA- positive S. saprophyticus with diverse origin carrying at least three different types of staphylococcal cassette chromosome mec (SCC mec ) are described here.     

Survey of Staphylococcus isolates among hospital personnel, environment and their antibiogram with special emphasis on methicillin resistance

The objective of this study was to find the prevalence of Staphylococcus spp. carriage among hospital personnel and hospital environment and their antibiogram with special emphasis on methicillin resistance. A total of 205 samples from hospital personnel and environment were collected from casualty, oncology and multidisciplinary cardiac unit ward of Kasturba Medical College Hospital, Manipal. Samples were collected using sterile cotton wool swabs and inoculated into brain heart infusion broth. Subcultures were done onto blood agar and MacConkey's agar. Isolates were identified by standard methods up to species level. Antimicrobial susceptibility test was performed according to standardized disc diffusion Kirby-Bauer method. Each of the isolates was screened for methicillin resistance using oxacillin disc on Mueller Hinton agar plate followed by MIC for methicillin and cefoxitin susceptibility test by disc diffusion method. Sixty five out of 205 strains (31.7%) were Staphylococcus spp. and all of them were coagulase negative. Most of the strains belonged to S.epidermidis 49.23% (32/65) followed by S. saprophyticus 26.15% (17/65). Maximum isolates of S.epidermidis were from anterior nares 28.12% (9/32 strains of S.epidermidis). Highest number of methicillin resistant coagulase negative strains (3/9, 33.33%) were isolated from stethoscope of multidisciplinary cardiac unit ward followed by carriers in the anterior nares (2/9, 22.22%). Methicillin resistant coagulase negative staphylococci are prevalent in anterior nares of hospital personnel and in the hospital environment thereby providing a definite source for hospital acquired infection. All isolates were sensitive to vancomycin, ciprofloxacin and amikacin.     

糖尿病足感染中耐甲氧西林表皮葡萄球菌SCCmec分型临床研究

目的 研究分离于糖尿病足感染(DFI)溃疡创面的耐甲氧西林表皮葡萄球菌(MRSE)的SCCmec分型特点及不同型别的耐药特征.方法 2008年1月-2010年6月从天津某医院390例DFI溃疡中分离出70株表皮葡萄球菌,提取细菌的DNA,用聚合酶链反应(PCR)方法检测mecA基因,并对MRSE进行SCCmec分型;采...     

Accuracy of cefoxitin disk testing for characterization of oxacillin resistance mediated by penicillin-binding protein 2a in coagulase-negative staphylococci

The Clinical and Laboratory Standards Institute (CLSI) proposed, beginning in 2004, the use of cefoxitin disks to predict resistance mediated by the mecA gene in all species of coagulase-negative staphylococci (CoNS). The aim of this work was to evaluate the efficiency of the cefoxitin disk and of oxacillin-salt agar screening (MHOX) to characterize the oxacillin resistance mediated by the mecA gene in CoNS. One hundred seven CoNS isolates from different clinical samples were studied. Detection of the mecA gene by PCR was considered the "gold standard." The susceptibility to oxacillin and cefoxitin was detected by the disk diffusion and agar dilution tests, as described by the CLSI. MHOX was also performed with 6 microg/ml of oxacillin and 4% NaCl. The sensitivities of the oxacillin and cefoxitin disks for all CoNS species were 88% and 80%, respectively, whereas the specificities were 63% and 100%, respectively. The sensitivities of the agar dilution test for oxacillin and cefoxitin (for proposed breakpoints of > or =4 microg/ml for resistance and < or =2 microg/ml for susceptibility) were 90% and 85%, respectively, whereas the specificities were 76% and 98%, respectively. MHOX showed a sensitivity of 90% and a specificity of 95% for all CoNS species. Both the MHOX and the cefoxitin disk results indicate that these are appropriate methods for the evaluation of oxacillin resistance mediated by the mecA gene in all CoNS species.     

Studies on clinical isolates of coagulase-negative staphylococci resistant to methicillin. Evidence of cross-resistance between methicillin and cephalothin

The in vitro susceptibility to cephalothin and cefuroxime of 195 isolates of methicillin-resistant coagulase-negative staphylococci was determined by the agar-diffusion test, using 7.5% NaCl-supplemented agar. The distribution of the inhibition zone diameters for isolates of S. epidermidis (S. biotype 1) as well as for S. haemolyticus (S. biotype 4) was trimodal. While 4% of the isolates were found susceptible to cefuroxime, 39% of the S. epidermidis/S. hominis (S. biotype 1) isolates and 34% of the S. haemolyticus (S. biotype 4) isolates were found susceptible to cephalothin by this method. Eight of these isolates (six S. epidermidis, two S. haemolyticus) were selected for susceptibility testing by the tube-dilution method, together with four isolates (three S. haemolyticus, one S. epidermidis) found resistant to cephalothin by the agar-diffusion test. The first-mentioned isolates were all found susceptible to cephalothin with MICs less than or equal to 2 micrograms/l, while the last-named all were resistant with MICs greater than or equal to 16 micrograms/ml. Population analyses revealed sub-populations of highly resistant bacteria in all methicillin-resistant isolates of S. epidermidis (S. biotype 1), as well as in all isolates of S. haemolyticus (S. biotype 4). We thus concluded that methicillin-resistance in isolates of coagulase-negative staphylococci implies resistance to cephalosporins and that the difference between S. epidermidis and S. haemolyticus as regards cephalosporin-susceptibility is quantitative and not qualitative. Eighty-nine per cent of the 195 methicillin-resistant isolates in this study were resistant to penicillin and at least one more antibiotic. We therefore think that resistance to penicillin and one or more non-beta-lactam antibiotics strongly suggests methicillin-resistance and that such isolates should be further tested on hypertonic media.     

Evaluation of three techniques for detection of low-level methicillin-resistant Staphylococcus aureus (MRSA): a disk diffusion method with cefoxitin and moxalactam,the Vitek 2 system,and the MRSA-screen latex agglutination test

Very-low-level methicillin-resistant Staphylococcus aureus (MRSA), or class 1 MRSA, is often misdiagnosed as methicillin-susceptible S. aureus (MSSA). We evaluated the performances of three methods for detection of low-level methicillin resistance: the disk diffusion method using the cephamycin antibiotics cefoxitin and moxalactam, the Vitek 2 system (bioMérieux), and the MRSA-screen test (Denka). Detection of the mecA gene by PCR was considered to be the "gold standard." We also determined the sensitivity of the oxacillin disk diffusion method with 5- and 1-microg disks and that of the Oxascreen agar assay with 6 mg of oxacillin liter(-1) for detection of MRSA. We compared the distributions of MICs of oxacillin and cefoxitin by the E-test (AB Biodisk), and those of moxalactam by dilutions in agar, for MRSA and MSSA isolates. The 152 clinical isolates of S. aureus studied were divided into 69 MSSA (mecA-negative) and 83 MRSA (mecA-positive) isolates, including 63 heterogeneous isolates and 26 class 1 isolates (low-level resistance). The cefoxitin and moxalactam disk diffusion tests detected 100% of all the MRSA classes: cefoxitin inhibition zone diameters were <27 mm, and moxalactam inhibition zone diameters were <24 mm. The Vitek 2 system and the MRSA-screen test detected 94 and 97.6% of all MRSA isolates, respectively. The sensitivities of the 5- and 1-microg oxacillin disks were 95.2 and 96.4%, respectively, whereas that of the Oxascreen agar screen assay was 94%. All of the tests except the 1-microg oxacillin disk test were 100% specific. For the class 1 MRSA isolates, the sensitivity of the Vitek 2 test was 92.3%, whereas those of the MRSA-screen test and the disk diffusion method with cefoxitin and moxalactam were 100%. Therefore, the cefoxitin and moxalactam disk diffusion methods were the best-performing tests for routine detection of all classes of MRSA.     

In vitro activities of eight antibiotics against methicillin-resistant S. aureus and S. epidermidis strains isolated in Korea

Staphylococcus aureus and Staphylococcus epidermidis strains isolated at eight large medical centers in Korea were examined for methicillin resistance and resistance to eight other antibiotics; cefazolin, cefamandole, cefuroxime, cefoxitin, cefotaxime, moxalactam, penicillin G and vancomycin. Methicillin resistance was found in 296 of 1225 strains (24.2%) of S. aureus and 126 of 348 strains (36.2%) of S. epidermidis. Methicillinresistant strains were isolated from all sources with the frequency of isolation ranging from 11% to 60%. From pleural effusion, throat swab and blood, methicillin-resistant strains of S. aureus were more frequently isolated with statistical significance (Chi-squared test, 95% confidence). Almost all of Methicillin-resistant S. aureus (MRSA) and S. epidermidis (MRSE) strains were multiply resistant to one or more tested eight antibiotics. However only 7(2.4%) of 296 MRSA strains and 2(1.6%) of 126 MRSE strains were resistant to vancomycin. Vancomycin was the most effective antibiotic against staphylococcal isolates as well as MRSA and MRSE.     

Correlation of oxacillin MIC with mecA gene carriage in coagulase-negative staphylococci

The National Committee for Clinical Laboratory Standards has recently changed the oxacillin breakpoint from >/=4 mg/liter to >/=0. 5 mg/liter to detect methicillin-resistant coagulase-negative staphylococci (CoNS) because the previous breakpoint lacked sensitivity. To determine the correlation between the new oxacillin breakpoint and the presence of the mecA gene, 493 CoNS of 11 species were tested. The presence of the mecA gene was determined by PCR, and oxacillin susceptibility was determined by the agar dilution method with Mueller-Hinton agar containing 2% NaCl and oxacillin (0. 125 to 4.0 mg/liter). The new breakpoint correctly classified all CoNS strains with mecA as methicillin resistant and strains of Staphylococcus epidermidis, S. haemolyticus, and S. hominis without mecA as methicillin susceptible. The breakpoint of >/=0.5 mg/liter was not specific for S. cohnii, S. lugdunensis, S. saprophyticus, S. warneri, and S. xylosus, in that it categorized 70 of 74 strains of these species without mecA (94.6%) as methicillin resistant. The results of this study indicate that the new oxacillin breakpoint accurately identifies strains of CoNS with mecA but is not specific for strains of certain species of CoNS without mecA.     

Characterization of SCCmec elements in methicillin-resistant Staphylococcus epidermidis isolated from blood cultures from neonates during three decades

Staphylococcus epidermidis is a major cause of nosocomial infections in immunocompromised patients and the predominant pathogen in catheter-related infections and bloodstream infections. Approximately 70-80% of S. epidermidis carry the mecA gene encoding methicillin resistance. The mecA gene is located on a mobile genetic element, the staphylococcal cassette chromosome mec (SCCmec). The aim of this study was to characterize the SCCmec elements as well as the adjacent arginine catabolic mobile element (ACME) in 30 clinical blood isolates of mecA positive S. epidermidis obtained from neonates and collected over a period of three decades. The ccr and mec gene complexes were identified using PCR. The SCCmec elements were found among 29/30 isolates and 13 different combinations of ccr gene complexes and mec gene complexes were identified. Staphylococcus epidermidis regularly carried multiple copies of ccr gene complexes, but only one class of mec gene complex. Three isolates could be assigned the SCCmec type III (3A). The combinations of ccr gene complexes and the mec gene complexes differed among the three decades. The most frequent combination was class B mec in combination with ccr1 and ccr2. Staphylococcus epidermidis may constitute a large reservoir for SCCmec elements, and frequent exchange of mobile genetic elements between staphylococcal species may explain the emergence of new MRSA strains.     

A study of coagulase-negative staphylococci isolated from clinically significant infections at an Australian teaching hospital

Some 151 isolates of coagulase-negative staphylococci isolated from patients at an Australian teaching hospital were characterized by biochemical analysis, antibiotic sensitivity patterns and slime production. S. epidermidis was the predominant species (64%) isolated from clinically significant infections, and all S. epidermidis isolates from true bacteremias produced slime. Forty-nine per cent were resistant to methicillin and 61% to gentamicin. S. haemolyticus isolates from clinically significant infections also showed antibiotic resistance and 80% were resistant to more than five antibiotics. The importance of coagulase-negative staphylococci as pathogens in this large teaching hospital was confirmed.     

Antimicrobial susceptibility of Bacteroides fragilis group isolates in Europe

Objective  To evaluate the activity of old and newer antianaerobic drugs against clinical isolates of Bacteroides fragilis group strains from different parts of Europe.
Methods  Bacteroides fragilis group isolates from 37 laboratories in 19 countries were biochemically characterized. The MICs of seven antimicrobial agents were determined by the agar dilution method as recommended by the NCCLS. Production of beta-lactamase was detected by nitrocefin.
Results  There were 1284 B. fragilis group isolates included in the study. Abdominal infections and wounds were the most common sources of isolation and B. fragilis was the dominating species. Ninety-nine percent of the strains were resistant to ampicillin (breakpoint 2 mg/L), 6% to cefoxitin (64 mg/L), 15% to clindamycin (8 mg/L) and 9% to moxifloxacin (8 mg/L). Less than 1% were resistant to imipenem (16 mg/L), piperacillin-tazobactam (128 mg/L) and metronidazole (32 mg/L). Ninety-six percent of the isolates were beta-lactamase producers.
Conclusions  Antimicrobial resistance among the B. fragilis group is increasing.     

Evaluation of laboratory tests for detection of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

Few studies evaluating susceptibility testing of methicillin-resistant staphylococci have included isolates of Staphylococcus epidermidis, a known pathogen in many types of serious infections. We tested 175 S. epidermidis and 95 Staphylococcus aureus isolates to determine the most sensitive procedures for detecting methicillin-resistant staphylococci. Reference procedures included agar dilution with methicillin and 4% NaCl in the agar and broth microdilution with methicillin and 2% NaCl in cation-supplemented Mueller-Hinton broth. After 24 h of incubation, the results from both methods correlated well and were within 1 log2 dilution for all isolates tested. Only one-half of all resistant isolates (92 of 183) were detected at 18 h by using the standard disk diffusion technique with 5-micrograms methicillin disks, and even fewer were detected with 10-micrograms methicillin disks and newly recommended zone-size criteria. However, the standard disk diffusion method with 4% NaCl in the agar increased the sensitivity and specificity for identification of the proper phenotype to greater than 92%. The spread plate and new spot techniques, both using agar with 4% NaCl, were also sensitive methods. Of 47 S. epidermidis isolates tested against oxacillin, 6 (13%) were oxacillin susceptible but methicillin resistant. Two automated systems, the Automicrobic system (Vitek Systems) and MicroScan (American MicroScan), as well as two broth screening systems available from Remel and Austin Biological Laboratories, failed to detect several resistant isolates, depending on the species.     

In vitro activity of tedizolid and linezolid against <Emphasis Type="Italic">Staphylococcus epidermidis</Emphasis> isolated from prosthetic joint infections

Prosthetic joint infections (PJIs) are rare but long-lasting and are serious complications without any spontaneous resolution, requiring additional surgery and long-term treatment with antibiotics. Staphylococci are the most important aetiological agents of PJIs, and among the coagulase-negative staphylococci Staphylococcus epidermidis is the most common. However, S. epidermidis often displays multidrug resistance (MDR), demanding additional treatment options. The objective was to examine the effectiveness of tedizolid and linezolid against S. epidermidis isolated from PJIs. The standard antibiotic susceptibility pattern of S. epidermidis (n?=?183) obtained from PJIs was determined by disc diffusion test, and MIC was determined by Etest for tedizolid, linezolid, and vancomycin. Tedizolid displayed MIC values ranging from 0.094 to 0.5 mg/L (MIC₅₀: 0.19 mg/L, MIC₉₀: 0.38 mg/L), linezolid MIC values ranging from 0.25 to 2 mg/L (MIC₅₀: 0.75 mg/L, MIC₉₀: 1 mg/L), and vancomycin MIC values ranging from 0.5 to 3 mg/L (MIC₅₀ and MIC₉₀ both 2 mg/L). According to the disc diffusion test, 153/183 (84%) isolates were resistant to ≥3 antibiotic groups, indicating MDR. In conclusion, S. epidermidis isolates from PJIs were fully susceptible, and the MIC₅₀ and MIC₉₀ values for tedizolid were two- to four-fold dilution steps lower compared with linezolid. Tedizolid is not approved, and there are no reports of long-term treatment, but it may display better tolerability and fewer adverse effects than linezolid; it thus could be a possible treatment option for PJIs, alone or in combination with rifampicin.     

Evaluation of phenotypic and molecular methods for detection of oxacillin resistance in members of the Staphylococcus sciuri group

In this paper we report on an experimental evaluation of phenotypic and molecular methods as means for the detection of oxacillin resistance in members of the Staphylococcus sciuri group. A total of 109 S. sciuri group member isolates (92 S. sciuri isolates, 9 S. lentus isolates, and 8 S. vitulinus isolates) were tested by the disk diffusion method, the agar dilution method, the oxacillin salt-agar screening method, slide latex agglutination for PBP 2a, and PCR assay for mecA as the reference method. The mecA gene was detected in 29 S. sciuri isolates, and the true-positive and true-negative results of the other tests were defined on the basis of the presence or the absence of the mecA gene. For the different methods evaluated, the sensitivities and specificities were as follows: for the disk diffusion test with a 1-microg oxacillin disk, 100% and 55.9%, respectively; for the disk diffusion test with a 30-mug cefoxitin disk, 93.5% and 100%, respectively; for the agar dilution method, 100% and 50%, respectively; for the oxacillin salt-agar screen test (with 6 microg of oxacillin per ml and 4% NaCl) 100% and 100%, respectively; and for the slide latex agglutination test for PBP 2a, 100% and 100%, respectively. The disk diffusion test with various beta-lactam antibiotics was performed to evaluate their use for the prediction of oxacillin resistance. The results indicate that meropenem, cefazolin, cefamandole, cefuroxime, cefotetan, cefoperazone, cefotaxime, ceftriaxone, moxalactam, cefaclor, and cefprozil may be used as surrogate markers of oxacillin resistance, although further studies of their use for the detection of oxacillin resistance are required.     

Mueller–Hinton agar is superior to PDM blood agar for detection of methicillin-resistant Staphylococcus aureus

The aim of this study was to compare the expression of oxacillin resistance in methicillin-resistant Staphylococcus aureus (MRSA) on Paper Disc Method agar supplemented with 5% defibrinated blood (PDM blood agar) and Mueller–Hinton agar supplemented with 2% NaCl (MH NaCl agar) using different susceptibility tests. Fifty mecA- containing isolates of S. aureus, exhibiting 46 different pulsed-field gel electrophoresis patterns, were comparatively tested using the E test, the single disk diffusion test, and the multipoint inoculation technique, under various culture conditions. The E test incubated at 35 °C for 24 h (breakpoint of resistance ≥2.0 mg/L) detected 94% of the isolates on MH NaCl agar, compared with 28% for PDM blood agar ( P  < 0.05). The disk diffusion test (breakpoint ≤ 10 mm in diameter) under these incubation conditions detected resistance in 100% of the isolates on MH NaCl agar and in 80% of the isolates on PDM blood agar ( P  < 0.05). The multipoint technique (breakpoint ≥1 mg/L), applied at 35 °C for 24 h, detected 100% on MH NaCl agar and 46% on PDM blood agar ( P  < 0.05). Irrespective of the method of susceptibility testing evaluated, MH NaCl agar was superior to PDM blood agar for the detection of oxacillin resistance in mecA -containing S. aureus .