Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey

ABSTRACT: Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

RESUMO: Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

Genomic characterization, antimicrobial resistance profiles, enterotoxin, and biofilm production of methicillin-resistant Staphylococcus aureus isolated from clinical and animal products origins in Eastern Turkey / Caracterização genômica perfis de resistência antimicrobiana, produção de enterotoxinas e biofilme Staphylococcus aureus resistente à meticilina isolado de origem clínica e de produtos de origem animal no leste da Turquia

Staphylococcus aureus is an opportunistic and ubiquitous pathogen found in the skin, nares, and mucosal membranes of mammals. Increasing resistance to antimicrobials including methicillin has become an important public concern. One hundred and eight (108) S. aureus strains isolated from a total of 572 clinical and animal products samples, were investigated for their biofilm capability, methicillin resistance, enterotoxin genes, and genetic diversity. Although only one strain isolated from raw retail was found as a strong biofilm producer, the percentage of antimicrobial resistance pattern was relatively higher. 17.59% of S. aureus strains tested in this study were resistant to cefoxitin and identified as methicillin-resistant S. aureus (MRSA) isolates. mecA and mecC harboring S. aureus strains were detected at a rate of 2.79% and 0.93%, respectively. In addition, staphylococcal enterotoxin genes including Sea, Seb, Sec, and Sed genes were found to be 18.5%, 32.4%, 6.5% and 3.7%, respectively. The phylogenetic relationship among the isolates showed relationship between joint calf and cow milk isolates. Multi locus sequence typing (MLST) revealed three different sequence types (STs) including ST84, ST829, and ST6238. These findings highlight the development and spread of MRSA strains with zoonotic potential in animals and the food chain throughout the world.

Staphylococcus aureus é um patógeno dúctil e ubíquo encontrado na pele, narinas e membranas mucosas de mamíferos. O aumento da resistência aos antimicrobianos, incluindo a meticilina, tornou-se uma importante preocupação pública. Cento e oito (108) cepas de S. aureus isoladas de um total de 572 amostras clínicas e de produtos animais foram investigadas por sua capacidade de biofilme, resistência à meticilina, genes de enterotoxinas e diversidade genética. Embora apenas uma cepa isolada do cru tenha sido encontrada como forte produtora de biofilme, a porcentagem do padrão de resistência antimicrobiana foi relativamente maior. Parte das cepas (17,59%) de S. aureus testadas neste estudo eram resistentes à cefoxitina e identificadas como isolados de MRSA. mecA e mecC abrigando cepas de S. aureus foram detectados a uma taxa de 2,79% e 0,93%, respectivamente. Além disso, verificou-se que os genes da enterotoxina estafilocócica, incluindo os genes Sea, Seb, Sec e Sed, eram 18,5%, 32,4%, 6,5% e 3,7%, respectivamente. A relação filogenética entre os isolados mostrou relação entre os isolados de bezerro e leite de vaca. A tipagem de sequência multiloco (MLST) revelou três tipos de sequência diferentes (STs), incluindo ST84, ST829 e ST6238. Essas descobertas destacam o desenvolvimento e a disseminação de cepas de MRSA com potencial zoonótico em animais e na cadeia alimentar em todo o mundo.

Staphylococcal cassette chromosome mec elements from Staphylococcus intermedius group (SIG) isolates from dogs in a center for veterinary diagnostics in Brazil / Cassetes cromossômicos estafilocócicos mec de isolados do grupo Staphylococcus intermedius (GSI) de cães em um centro veterinário de diagnósticos no Brasil

Methicillin resistance in the Staphylococcus intermedius group (SIG) has emerged in small animal practice. Methicillin-resistant SIG (MRSIG) members have been implicated as causes of infections in both companion animals and humans. Staphylococcal cassette chromosome mec (SCCmec) elements carry the mecA/C genes, which encode for the transpeptidase PBP2a (PBP2’) responsible for β-lactam antibiotic resistance in staphylococci. This study examined the SCCmec types of MRSIG isolates from different clinical specimens of dogs that exhibited methicillin MIC ≥ 0.5 μg/mL by an automated identification and susceptibility system in a Center for Veterinary Diagnostics in São Paulo, Brazil. Susceptibility to methicillin was determined by broth microdilution testing, and Oxoid® M.I.C.Evaluator® strips. PBP2a production was detected using a latex agglutination assay. SCCmec typing was performed according to the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) guidelines. SCCmec type II (2A), SCCmec type III (3A), composite SCC structures consisting of a class A mec gene complex in addition to multiple ccr gene complexes, and non-typable SCCmec elements were reported in these MRSIG isolates. SCCmec type variants differing from those so far acknowledged by IWG-SCC were found, indicating new rearrangements in the genetic context of mecA in these canine MRSIG isolates.

A resistência à meticilina no grupo Staphylococcus intermedius (GSI) tem aumentado na clínica de pequenos animais. Membros GSI resistentes à meticilina (GSIRM) têm sido causas de infecções tanto em animais de companhia e humanos. Cassetes cromossômicos estafilocócicos mec (SCCmec) carregam os genes mecA/C, que codificam a transpeptidase PBP2a (PBP2’) responsável pela resistência aos antibióticos β-lactâmicos em estafilococos. Nosso objetivo foi investigar os elementos SCCmec de GSIRM isolados de diferentes amostras clínicas de cães que exibiram CIM de meticilina ≥ 0,5 μg/mL por meio de um sistema automatizado em um Centro Veterinário de Diagnósticos em São Paulo, Brasil. A sensibilidade à meticilina foi determinada por meio do teste de microdiluição em caldo e fitas Oxoid® M.I.C.Evaluator®. A produção de PBP2a foi detectada usando um ensaio de aglutinação de látex. A tipagem dos elementos SCCmec foi realizada de acordo com as diretrizes do International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). SCCmec tipo II (2A), SCCmec tipo III (3A), SCC compostos de um complexo mec de classe A com múltiplos complexos ccr, e elementos SCCmec não tipáveis foram encontrados nesses isolados GSIRM. Variantes que diferem dos elementos SCCmec reconhecidos até o momento pelo IWG-SCC foram encontradas, indicando novos rearranjos no contexto genético de mecA nesses isolados GSIRM caninos.

Staphylococcal cassette chromosome mec elements from Staphylococcus intermedius group (SIG) isolates from dogs in a center for veterinary diagnostics in Brazil / Cassetes cromossômicos estafilocócicos mec de isolados do grupo Staphylococcus intermedius (GSI) de cães em um centro veterinário de diagnósticos no Brasil

Methicillin resistance in the Staphylococcus intermedius group (SIG) has emerged in small animal practice. Methicillin-resistant SIG (MRSIG) members have been implicated as causes of infections in both companion animals and humans. Staphylococcal cassette chromosome mec (SCCmec) elements carry the mecA/C genes, which encode for the transpeptidase PBP2a (PBP2) responsible for β-lactam antibiotic resistance in staphylococci. This study examined the SCCmec types of MRSIG isolates from different clinical specimens of dogs that exhibited methicillin MIC ≥ 0.5 μg/mL by an automated identification and susceptibility system in a Center for Veterinary Diagnostics in São Paulo, Brazil. Susceptibility to methicillin was determined by broth microdilution testing, and Oxoid® M.I.C.Evaluator® strips. PBP2a production was detected using a latex agglutination assay. SCCmec typing was performed according to the International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC) guidelines. SCCmec type II (2A), SCCmec type III (3A), composite SCC structures consisting of a class A mec gene complex in addition to multiple ccr gene complexes, and non-typable SCCmec elements were reported in these MRSIG isolates. SCCmec type variants differing from those so far acknowledged by IWG-SCC were found, indicating new rearrangements in the genetic context of mecA in these canine MRSIG isolates.(AU)

A resistência à meticilina no grupo Staphylococcus intermedius (GSI) tem aumentado na clínica de pequenos animais. Membros GSI resistentes à meticilina (GSIRM) têm sido causas de infecções tanto em animais de companhia e humanos. Cassetes cromossômicos estafilocócicos mec (SCCmec) carregam os genes mecA/C, que codificam a transpeptidase PBP2a (PBP2) responsável pela resistência aos antibióticos β-lactâmicos em estafilococos. Nosso objetivo foi investigar os elementos SCCmec de GSIRM isolados de diferentes amostras clínicas de cães que exibiram CIM de meticilina ≥ 0,5 μg/mL por meio de um sistema automatizado em um Centro Veterinário de Diagnósticos em São Paulo, Brasil. A sensibilidade à meticilina foi determinada por meio do teste de microdiluição em caldo e fitas Oxoid® M.I.C.Evaluator®. A produção de PBP2a foi detectada usando um ensaio de aglutinação de látex. A tipagem dos elementos SCCmec foi realizada de acordo com as diretrizes do International Working Group on the Classification of Staphylococcal Cassette Chromosome Elements (IWG-SCC). SCCmec tipo II (2A), SCCmec tipo III (3A), SCC compostos de um complexo mec de classe A com múltiplos complexos ccr, e elementos SCCmec não tipáveis foram encontrados nesses isolados GSIRM. Variantes que diferem dos elementos SCCmec reconhecidos até o momento pelo IWG-SCC foram encontradas, indicando novos rearranjos no contexto genético de mecA nesses isolados GSIRM caninos.(AU)

Investigation of methicillin resistance and Panton-Valentine leukocidin in Staphylococci isolated from bovine mastitis

Background: Mastitis, which is inflammation of the mammary gland, is among the most important diseases in dairy herds resulting in reductions of milk yield and milk quality. Although several groups of microorganisms have been reported as etiological agents of mastitis, Staphylococci are the most frequently isolated bacteria from bovine mastitic milk samples. The aim of this study was to isolate the Staphylococcus species from bovine mastitis, investigate the mecA ve pvl genes in isolated species by PCR and determine the antibiotic resistance of methicillin resistant strains to some antibiotics commonly used in veterinary field. Materials, Methods & Results: In the present study, 972 half-udder milk samples (n = 757 CMT positive, n = 215 CMT negative) were used from 251 lactating cows from 34 different enterprises located center town and villages of Õdemi, zmir. Ten microliters of each milk sample was inoculated onto Columbia blood agar, containing 7% of sheep blood and incubated under aerobic conditions for 24-48 h at 37C. The certain identification of Staphylococcus isolates was achieved using Crystal Identification Systems Gram-Positive ID kit. Bacterial DNAs were extracted from all strains using boiling method and strains were screened for the presence of 16S rDNA, mecA and pvl genes by PCR. The antimicrobial resistance of MRS species was determined by using disc [...](AU)

Investigation of methicillin resistance and Panton-Valentine leukocidin in Staphylococci isolated from bovine mastitis

Background: Mastitis, which is inflammation of the mammary gland, is among the most important diseases in dairy herds resulting in reductions of milk yield and milk quality. Although several groups of microorganisms have been reported as etiological agents of mastitis, Staphylococci are the most frequently isolated bacteria from bovine mastitic milk samples. The aim of this study was to isolate the Staphylococcus species from bovine mastitis, investigate the mecA ve pvl genes in isolated species by PCR and determine the antibiotic resistance of methicillin resistant strains to some antibiotics commonly used in veterinary field. Materials, Methods & Results: In the present study, 972 half-udder milk samples (n = 757 CMT positive, n = 215 CMT negative) were used from 251 lactating cows from 34 different enterprises located center town and villages of Õdemi, zmir. Ten microliters of each milk sample was inoculated onto Columbia blood agar, containing 7% of sheep blood and incubated under aerobic conditions for 24-48 h at 37C. The certain identification of Staphylococcus isolates was achieved using Crystal Identification Systems Gram-Positive ID kit. Bacterial DNAs were extracted from all strains using boiling method and strains were screened for the presence of 16S rDNA, mecA and pvl genes by PCR. The antimicrobial resistance of MRS species was determined by using disc [...]

OCORRÊNCIA, IDENTIFICAÇÃO DE FATORES PRESDISPONENTES E CARACTERIZAÇÃO DE Staphylococcus sp. RESISTENTES À METICILINA EM CÃES

Staphylococcus sp. são importantes tanto na medicina humana como na medicina veterinária. Embora possa ser encontrado na pele e mucosas de animais portadores assintomáticos, está relacionado a graves quadros de piodermite e otites em cães. A espécie que mais acomete esses animais é o S. pseudintermedius, que também já vem sendo encontrado em infecções humanas, e em menor frequência, mas não menos importante, S. schleiferi. Com o estreitamento do convívio entre animais e os seres humanos, sabendo do potencial zoonótico e antropozoonótico desses microrganismos e crescente aumento de bactérias multirresistentes, traz uma grande necessidade de aprofundar estudos sobre o tema e passar a ter uma nova leitura sobre a saúde animal influenciando na saúde humana (conceito One health). Desta forma o presente estudo teve como objetivo caracterizar amostras de Staphylococcus sp. isoladas de cães com infecções tópicas bacterianas (otite e dermatite) quanto a sua distribuição de espécies, resistência aos antimicrobianos, especialmente a meticilina e alguns fatores de virulência. Com esse objetivo amostras de cem animais com dermatopatias foram coletadas com auxílio de swab estéril de lesões bacterianas de pele e conduto auditivo, possibilitando o isolamento de 350 UFC. A espécie mais isolada foi S. pseudintermedius (37,7%), seguido S. scheleiferi (17%). No teste de sensibilidade aos antimicrobianos, as penicilinas, macrolídeos e quinolonas apresentaram maior percentual de resistência nos MSS. Nos MRS foram as lincosaminas, quinolonas e macrólideos. O gene mecA foi detectado em 21,7% das amostras (31% dos cães). A presença de genes de que codificam proteínas de superfície (sps) foi avaliada nos S. pseudintermedius, sendo a proteína spsA a mais encontrada em MSSP (96%) e spsR nos MRSP (95%). Dentre os possíveis fatores predisponentes correlacionados a incidência de Staphylococcus sp. resistentes à meticilina, a idade e o local onde dormem mostraram relevância estatística. A partir desses dados conclui-se que há necessidade de monitorar a resistência a meticilina nos cães, pois sua presença e risco são eminentes, ainda mais com estreitamento do convívio entre animais e seus tutores. E S. pseudintermedius, dentre os Staphylococcus, permanece sendo o mais isolado nas afecções de pele.

Staphylococcus sp. are importante in both human and veterinary medicine. Although it can be found in the skin and mucous membranes of animals, making them only carriers, it is realted to severe pyoderma and otitis in dogs. The species that most affects these animals is S. pseudintermedius, which has also been found in human infections, and less frequently, but not least, S. scheleiferi. With the closer relationship between animals and humans, knowing the zoonotic and anthropozoonotic potencial of these microoganisms and the growing increase of multiresistant bacteria, brings a great need to deepen stidies on the subject and to have a new Reading on animal health influencing the human health (One health concept). Thus, the presente study aims to characterize samples of Staphylococcus sp. resistant to methicillin isolates from dogs with topical bacterial infections (otitis and dermatites) for species diversity, Antimicrobial susceptibility profile, virulence genes and ocurrence of predisposing factors. To that end samples of one hundred animals with dermatopathies were collected with the aid of sterile swab of bacterial skin lesions and ear canal, thus enable isolation of 350 UFC. The most isolated species was S. pseudintermedius (37,7%), followed by S. scheleiferi (17%). In the antimicrobial susceptibility profile, penicillinases, macrolides and quinolones showed a higher percentage of resistance in MSS. In the MRS were lincosamines, quinolones and macrolides. The mecA gene was detected in 21,7% of the samples (31% of the dogs). And the presence of adherence genes (sps) in S. pseudintermedius was investigated, with spsA protein being the most found in MSSP (96%) and spsR in MRSP (95%). Among the possible predisposing factors correlated to the incidence of Staphylococcus sp. resistant to methicilin, age and the place of sleep showed statistical relevance. From these data, assumed that there is a need to monitor methicillin resistance in dogs, as its presence and risk are iminente, especially with the narrowing of the interaction between animals their guardians. And S. pseudintermedius remains the most isolated in skin disorders.

Methicillin-resistant Staphylococcus sp. colonizing health care workers of a cancer hospital

The aim of the study was to analyze epidemiological and microbiological aspects of oral colonization by methicillin-resistant Staphylococcus of health care workers in a cancer hospital. Interview and saliva sampling were performed with 149 health care workers. Antimicrobial resistance was determined by disk diffusion and minimum inhibitory concentration. Polymerase Chain Reaction, Internal Transcribed Spacer-Polymerase Chain Reaction and Pulsed Field Gel Electrophoresis were performed for genotypic characterization of methicillin-resistant Staphylococcus. Risk factors were determined by logistic regression. Methicillin-resistant Staphylococcus colonization prevalence was 19.5%, denture wearing (p = 0.03), habit of nail biting (p = 0.04) and preparation and administration of antimicrobial (p = 0.04) were risk factors identified. All methicillin-resistant Staphylococcus were S. epidermidis, 94.4% of them had mecA gene. Closely related and indistinguishable methicillin-resistant S. epidermidis were detected. These results highlight that HCWs which have contact with patient at high risk for developing infections were identified as colonized by MRSE in the oral cavity, reinforcing this cavity as a reservoir of these bacteria and the risk to themselves and patients safety, because these microorganisms may be spread by coughing and talking.

Detection of femA and mecA genes in Staphylococcus aureus isolated from bovine raw milk using duplex PCR and determination of the minimum inhibitory concentration of the isolates / Uso de PCR Duplex para detecção dos genes femA e mecA e determinação da concentração inibitória mínima (CIM) em Staphylococcus aureus isolados de leite cru

This study identified Staphylococcus aureus and the methicillin-resistant (MRSA) strains in raw milk samples using molecular methods, and their minimum inhibitory concentrations (MIC) were evaluated. Two hundred fifty-one samples of raw milk samples from bulk tanks, collected in the state of Minas Gerais, Brazil were analyzed using polymerase chain reaction (PCR) to investigate the presence of femA gene in 278 S.aureus isolates. Furthermore, MRSA strains were investigated for detecting mecA gene. All of 278 isolates contained the femA gene (132 bp), and in 11 samples (3.95 %) the mecA gene (533 bp) was identified. The minimum inhibitory concentration against S. aureus was determined using enrofloxacin, trimethoprim-sulfamethoxazole, gentamicin, azithromycin, tetracycline, cephalothin, amoxicillin, penicillin G and lincomycin. Enrofloxacin and trimethoprim-sulfamethoxazole showed to be the most effective inhibitors, as this study identified 149 of 161 isolates (92.5 % of susceptibility) with MIC≤ 4 mg/ mL, and 55 of 66 isolates (83.3 % of susceptibility) with MIC≤ 2 mg/mL, for the first antimicrobial drug and the second one, respectively.(AU)

O objetivo deste trabalho foi de identificar Staphylococcus aureus e as linhagens resistentes à meticilina (MRSA) em amostras de leite cru bovino, por meio de métodos moleculares, e de determinar a concentração inibitória mínima (CIM) dos isolados. S. aureus pode apresentar linhagens resistentes à meticilina e a outros antimicrobianos utilizados na prática médico-veterinária. Foram analisadas 251 amostras de leite cru de tanques de expansão, procedentes de cinco mesorregiões produtoras de leite de Minas Gerais Brasil. Por meio de PCR foi identificada a presença do gene femA em 278 isolados de S. aureus; e as linhagens MRSA foram determinadas pela presença do gene mecA. Os ensaios de PCR foram positivos para gene femA em 100 % dos isolados testados; e entre essas amostras 11 (3,95 %) apresentaram produto amplificado de 533 pb correspondente ao gene mecA. Para analisar a susceptibilidade dos isolados foi realizado o teste de CIM para nove substâncias antimicrobianas. Foram identificados 149 isolados de S. aureus que apresentaram CIM≤ 4 µg/mL para enrofloxacina, com 92,54 % de susceptibilidade, e 55 isolados testados para sulfametoxazol+trimetoprima, apresentaram CIM ≤ 2 µg/mL com 83,33 % de susceptibilidade. Estes antimicrobianos foram considerados os mais eficazes.(AU)

Bactericidal antibiotic-phytochemical combinations against methicillin resistant Staphylococcus aureus

Methicillin resistant Staphylococcus aureus (MRSA) infection is a global concern nowadays. Due to its multi-drug resistant nature, treatment with conventional antibiotics does not assure desired clinical outcomes. Therefore, there is a need to find new compounds and/or alternative methods to get arsenal against the pathogen. Combination therapies using conventional antibiotics and phytochemicals fulfill both requirements. In this study, the efficacy of different phytochemicals in combination with selected antibiotics was tested against 12 strains of S. aureus (ATCC MRSA 43300, ATCC methicillin sensitive S. aureus or MSSA 29213 and 10 MRSA clinical strains collected from National University Hospital, Singapore). Out of the six phytochemicals used, tannic acid was synergistic with fusidic acid, minocycline, cefotaxime and rifampicin against most of strains tested and additive with ofloxacin and vancomycin. Quercetin showed synergism with minocycline, fusidic acid and rifampicin against most of the strains. Gallic acid ethyl ester showed additivity against all strains in combination with all antibiotics under investigation except with vancomycin where it showed indifference effect. Eugenol, menthone and caffeic acid showed indifference results against all strains in combination with all antibiotics. Interestingly, no antagonism was observed within these interactions. Based on the fractional inhibitory concentration indices, synergistic pairs were further examined by time-kill assays to confirm the accuracy and killing rate of the combinations over time. The two methods concurred with each other with 92% accuracy and the combinatory pairs were effective throughout the 24 hours of assay. The study suggests a possible incorporation of effective phytochemicals in combination therapies for MRSA infections.

Evaluation of different methods for detecting methicillin resistance in Staphylococcus aureus isolates in a university hospital located in the Northeast of Brazil

Many methods have been described for the detection of methicillin-resistant Staphylococcus aureus (MRSA), but the heterogeneous expression of methicillin resistance affects the reliability of these methods. The aim of the present study was to evaluate some methods for detecting methicillin resistance in Staphylococcus aureus isolates in a university hospital located in the Northeast of Brazil. Among the isolates, 15 were methicillin-susceptible and 45 were methicillin-resistant, including low-level heterogeneous resistance strains. Both the 30 µg-cefoxitin disk and PBP2a test had 100% sensibility/specificity and appear to be good options for the detection of MRSA in the clinical laboratory.