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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. RESUMO: 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.

Resumen Desde el inicio de la era antimicrobiana se han ido seleccionando gradualmente cepas de Staphylococcus aureus resistentes a antimicrobianos de amplio uso clínico. Es así como en 1960 se describen en Inglaterra las primeras cepas resistentes a meticilina, y algunos años después son informadas en hospitales de Chile. Actualmente, S. aureus resistente a penicilinas antiestafilocóccicas es endémico en los hospitales de nuestro país y del mundo, siendo responsable de una alta morbimortalidad. La resistencia es mediada habitualmente por la síntesis de una nueva transpeptidasa, denominada PBP2a o PBP2’ que posee menos afinidad por el β-lactámico, y es la que mantiene la síntesis de peptidoglicano en presencia del antimicrobiano. Esta nueva enzima se encuentra codificada en el gen mecA, a su vez inserto en un cassette cromosomal con estructura de isla genómica, de los cuales existen varios tipos y subtipos. La resistencia a meticilina se encuentra regulada, principalmente, por un mecanismo de inducción de la expresión del gen en presencia del β-lactámico, a través de un receptor de membrana y un represor de la expresión. Si bien se han descrito mecanismos generadores de resistencia a meticilina mec independientes, son categóricamente menos frecuentes.

Staphylococcus aureus resistente à meticilina (MRSA), outrora isolado especificamente em ambientes hospitalares, vem sendo identificado como causador de infecções cutâneas em pacientes da comunidade. Neste artigo, é relatado um caso do sul do Brasil com furunculose por CA-MRSA. O microrganismo isolado foi submetido a exames de PCR para o gene mecA e para o gene que codifica a leucocidina de Panton-Valentine. Esses exames permitiram a identificação genotípica do CA-MRSA.Methicillin-resistant Staphylococcus aureus (MRSA), particularly isolated at hospital setting, has been identified in cutaneous infections of community patients. This paper reports a case of furunculosis from the southern Brazil. Dermatologists must be attentive to this emergent etiological diagnosis. The isolated microorganism was subjected to PCR for gene mecA and to PCR for the gene that encodes the leukocidin of Panton-Valentine. These exams enabled genotypic identification of CA-MRSA.

PURPOSE: To evaluate different methods of oxacillin susceptibility testing of ocular isolates, considering polymerase chain reaction (PCR) as the 'gold standard', and to compare the in vitro susceptibility to oxacillin with that of other antimicrobials used in ophthalmologic practice. METHODS: The Vitek gram-positive identification card was used to identify ocular coagulase negative Staphylococcus species. The presence of the mecA gene was determined by the polymerase chain reaction assay with a combination of two primer sets (mecA and 16S rRNA) in a single region. Results were analyzed and compared with other oxacillin susceptibility methods: PBP2a detection by rapid slide latex agglutination test (SLA); oxacillin E-test; the Vitek automated gram-positive susceptibility card (GPS-105); the oxacillin salt agar screening test (OSAS) at a concentration of 6.0, 1.0 and 0.75 µg oxacillin per ml and the cefoxitin disk diffusion test (CDD). Automated susceptibility was also determined to other antimicrobial agents (fluoroquinolones, penicillin G, amoxicillin-ampicillin, cefazolin, ampicillin-sulbactam, erythromycin, clindamycin, gentamicin, tetracycline, trimethoprim-sulfamethoxazole, vancomycin and rifampin. RESULTS: Of the 69 CoNS isolates tested, 71% were mecA-positive and 29% mecA-negative. All methods tested had a statistically significant agreement with polymerase chain reaction. There was a tendency of positive polymerase chain reaction predomination among the S. epidermidis isolates in comparison to non-epidermidis isolates, although this was not statistically significant (78.3% vs. 56.5%; chi2= 2.54; P= 0.11). The oxacillin salt agar screening test (0.75 µg oxacillin/ml) showed the best performance, with 100% sensitivity and negative predictive value; 95% specificity and 98% positive predictive value. Using the E-test, the mecA-positive isolates were statistically significantly more resistant to ciprofloxacin, ofloxacin, gatifloxacin and moxifloxacin (P= 0.002; P= 0.008; P= 0.002 and P= 0.003, respectively). There was a statistically significant higher proportion of resistance of the coagulase negative Staphylococcus mecA-positives for: penicillin G, amoxicillin-ampicillin, cefazolin, ampicillin-sulbactam, erythromycin, clindamycin, gentamicin and tetracycline (POBJETIVOS: Avaliar os diferentes métodos de suscetibilidade à oxacillina, em isolados oculares, considerando a reação em cadeia da polimerase (PCR) como "padrão-ouro" e comparar a suscetibilidade in vitro para outros antimicrobianos de uso oftalmológico. MÉTODOS: O sistema automatizado Vitek foi utilizado para identificar as diferentes espécies de Staphylococcus coagulase negativo (SCoN). A presença do gene mecA foi determinado pela reação em cadeia da polimerase com a combinação de 2 "primer" sets (mecA e 16S rRNA) em uma única região. Estes resultados foram analisados e comparados com outros métodos de suscetibilidade à oxacilina: detecção da proteína PBP2a pelo teste de aglutinação em látex (SLA); E-test oxacilina; o sistema automatizado Vitek (GPS-105); o teste de triagem em ágar (OSAS) com oxacilina nas concentrações de 6,0, 1,0 e 0,75 µg oxacilina por ml e o teste de disco difusão com cefoxitina (CDD). A suscetibilidade automatizada foi obtida para os seguintes agentes antimicrobianos: fluorquinolonas, penicilina G, amoxicilina-ampicilina, cefazolina, ampicilina-sulbactam, eritromicina, clindamicina, gentamicina, tetraciclina, sulfametoxazol-trimetoprima, vancomicina e rifampicina. RESULTADOS: Dos 69 Staphylococcus coagulase negativo testados, 71% foram mecA-positivos e 29%, mecA-negativos. Todos os métodos testados apresentaram concordância estatisticamente significante com a reação em cadeia da polimerase. Houve tendência à predominância da positividade da reação em cadeia da polimerase entre os S. epidermidis comparado aos não-epidermidis, embora sem significância estatistica (78,3% vs. 56,5%; chi2= 2,54; p=0,11). O teste de triagem em ágar (0,75 µg oxacilina/ml) apresentou a melhor performance com resultados de: 100% de sensibilidade e valor preditivo negativo, 95% de especificidade e 98% de valor preditivo positivo. Os isolados mecA-positivos foram estatisticamente significativavos mais resistentes para ciprofloxacina, ofloxacina, gatifloxacina e moxifloxacina, no E-test (p=0,002; p=0,008; p=0,002 e p=0,003). Houve maior proporção estatisticamente significativa de resistência entre os Staphylococcus coagulase negativo mecA-positivos para: penicilina G, amoxicilina-ampicilina, cefazolina, ampicilina-sulbactam, eritromicina, clindamicina, gentamicina e tetraciclina. (p

PURPOSE: To evaluate the fluoroquinolone susceptibilities of ocular isolate coagulase-negative staphylococci (CoNS), identified at the Microbiology Laboratory - UNIFESP. DESIGN: Experimental laboratory investigation. METHODS: The minimum inhibitory concentrations (MICs) of 21 strains of methicillin-resistant coagulase-negative staphylococci (MRCoNS) and 22 methicillin-sensitive coagulase-negative staphylococci (MSCoNS) to ciprofloxacin, ofloxacin, gatifloxacin and moxifloxacin were determined, using the E-test method standardized by the Clinical and Laboratory Standards Institute (CLSI/NCCLS). RESULTS: The MIC90s (µg/ml) for the second generation of tested fluoroquinolones were higher than the fourth generation, especially for the methicillin-resistant coagulase-negative staphylococci group. CONCLUSION: Our results indicate that methicillin-sensitive coagulase-negative staphylococci are more susceptible to quinolones than are methicillin-resistant coagulase-negative staphylococci and that fourth generation fluoroquinolones appear to be more potent, affecting even coagulase-negative staphylococcal strains resistant to second generation fluoroquinolones.OBJETIVOS: Avaliar a suscetibilidade a fluorquinolonas dos Staphylococcus coagulase-negativo (SCoN) identificados no Laboratório de Microbiologia Ocular da Unifesp. MÉTODOS: Foi determinada a concentração inibitória mínima de 21 cepas de SCoN meticilina-resistentes e 22 meticilina-sensíveis para ciprofloxacina, ofloxacina, gatifloxacina e moxifloxacina, utilizando o E-test estandartizado pelo CLSI/NCCLS. RESULTADOS: Os MIC90 (µg/ml) de 43 SCoN isolados para fluorquinolonas de segunda geração foram maiores do que os de quarta geração, principalmente para o grupo dos meticilina-resistentes. CONCLUSÃO: Nossos resultados indicam que Staphylococcus coagulase-negativo meticilina-sensíveis são mais suscetíveis às quinolonas do que os Staphylococcus coagulase-negativo meticilina-resistentes, fluorquinolonas de quarta geração parecem ser mais potentes, cobrindo inclusive cepas de Staphylococcus coagulase-negativo resistentes à segunda geração de fluorquinolonas.

Staphylococcus aureus meticilino-resistente (MRSA) es un patógeno que ha emergido en las últimas cuatro décadas causando tanto infecciones nosocomiales como de la comunidad. La rápida y precisa detección de MRSA es relevante para guiar una apropiada terapia antibiótica y evitar la diseminación nosocomial de MRSA.En este trabajo se evaluó la eficiencia de métodos convencionales para la detección de meticilino-resistencia como difusión por discos, CIM en medio sólido, screening de oxacilina, y el nuevo test de aglutinación MRSA-Screen latex sobre 100 aislamientos de S. aureus, 79 mecA positivos y 21 mecA negativos. El test de aglutinación MRSA-Screen latex (Denka Seiken, Niigata, Japón) detecta la presencia de la PLP-2a, producto del gen mecA en cepas de S. aureus. La detección del gen mecA por PCR se utilizó como gold standard para comparar los resultados de los diferentes métodos. La sensibilidad y especificidad fueron 97 y 100 % para el método de difusión, 97 y 95 % para la CIM en medio sólido, 100 y 100 % para el screening de oxacilina y 100 y 100 % para MRSA-Screen latex. Todos los métodos presentaron alta sensibilidad y especificidad, pero el “MRSA-Screen latex” mostró la ventaja de poder brindar un resultado confiable, equivalente a la PCR, en sólo 15 minutos.Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen that has emerged over the last four decades, causing both nosocomial and community-acquired infections. Rapid and accurate detection of methicillin resistance in S. aureus is important for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of MRSA strains. We evaluated the efficiency of conventional methods for detection of methicillin resistance such as the disk diffusion, agar dilution, oxacillin agar screen test, and the latex agglutination test MRSA-Screen latex, in 100 isolates of S. aureus, 79 mecA positive and 21 mecA negative. The MRSA-Screen latex (Denka Seiken, Niigata, Japón), is a latex agglutination method that detects the presence of PLP-2a, product of mecA gene in S. aureus. The PCR of the mecA gene was used as the “gold standard” for the evaluation of the different methods tested. The percentages of sensitivity and specificity were as follows: disk difusión 97 and 100 %, agar dilution 97 and 95 %, oxacillin agar screen test 100 and 100 %, and MRSA-Screen latex, 100 and 100 %. All methods presented high sensitivity and specificity, but MRSA-Screen latex had the advantage of giving a reliable result, equivalent to PCR, in only 15 minutes.

En México como en otros países, Staphylococcus aureus es uno de los principales patógenos causantes de mastitis bovina. El tratamiento de esta enfermedad se realiza principalmente con antibióticos PTactámicos; sin embargo, en México existen estudios de Staphylococcus aureus resistente a este tipo de antibióticos, actualmente los mecanismos moleculares de Staphylococcus aureus resistente a meticilina no han sido ampliamente estudiados. En este trabajo se identificó Staphylococcus aureus aislados de bovinos mediante la detección del gen femB, utilizando ambos métodos, microbiológicos convencionales y moleculares. La resistencia a los antibióticos β-lactámicos fue determinada por pruebas de susceptibilidad antimicrobiana in vitro y por PCR de los genes blaZ, mecA y los genes reguladores mecR1 y mecI. Los resultados mostraron una mayor resistencia de Staphylococcus aureus a los antibióticos PTactámicos y a la eritromicina, solamente 38 (44,7%) de los aislamientos fueron positivos al gen blaZ. Por otro lado, cuatro Staphylococcus aureus fueron positivos al gen mecA considerados como SARM, los que por la detección de sus genes reguladores se agruparon de la siguiente forma: uno fue positivo a los genes mecI y mecR1 (grupo A), dos no presentaron el gen mecI ni el dominio PB del gen mecR1 (grupo C) y uno no mostró ningún gen regulador (grupo D). En conclusión, se identificó la presencia de Staphylococcus aureus resistente a la meticilina en unidades de producción lechera familiar en el estado de México, y su resistencia a los antibióticos β-lactámicos. Esta resistencia podría estar regulada por diferentes genotipos de los genes mecA, mecR1 y mecI presentes en Staphylococcus aureus resistente a la meticilina.

Community-associated methicillin-resistantStaphylococcus aureus(CA-MRSA) is an emerging public health problem worldwide. Severe invasive infections have been described, mostly associated with the presence of Panton-Valentine leukocidin (PVL). In Portugal limited information exists regarding CA-MRSA infections. In this study we describe the case of a previously healthy 12-year-old female, sport athlete, who presented to the hospital with acetabulofemoral septic arthritis, myositis, fasciitis, acetabulum osteomyelitis, and pneumonia. The MRSA isolated from blood and synovial fluid was PVL negative and staphylococcal enterotoxin type P (SEP) and type L (SEL) positive, with a vancomycin MIC of 1.0 mg/L and resistant to clindamycin and ciprofloxacin. The patient was submitted to multiple surgical drainages and started on vancomycin, rifampicin, and gentamycin. Due to persistence of fever and no microbiological clearance, linezolid was started with improvement. This is one of the few reported cases of severe invasive infection caused by CA-MRSA in Portugal, which was successfully treated with linezolid. In spite of the severity of infection, the MRSA isolate did not produce PVL.

Introducción: Staphylococcus aureus (SA) ha adoptado como nicho habitual en las narinas anteriores; sin embargo, recientemente se han reportado tasas de colonización del tracto genital de mujeres embarazadas entre 14% y 17.1%, lo que aumenta el interés respecto al riesgo asociado para el neonato y la madre en el posparto. Objetivo: Determinar la prevalencia de colonización nasal y vaginal por SA y los patrones de susceptibilidad a antibióticos de los aislamientos en una comunidad obstétrica de Cartagena, Colombia. Métodos: Se obtuvieron hisopados nasales y vaginales de las participantes y se sometieron a ensayos microbiológicos y moleculares. Se realizó seguimiento a la madre y al neonato durante cuatro semanas. Resultados: De 100 embarazadas participantes, 34 estuvieron colonizadas con SA; 29 solo en fosas nasales, tres solo vaginal y dos en ambos sitios. La colonización fue más común en fosas nasales que en vagina o en ambos sitios [29/34 (85.3%) vs 3/34 (8.8%) y 2/34 (5.9%); p

Introduction: The host niche for Staphylococcus aureus (SA) are the anterior nares; however, vaginal colonization rates between 14% and 17.1% in pregnant women have been recently reported, raising interest about the potential risk in postpartum women and in neonates from colonized mothers. Objectives: To determine the prevalence of nasal and vaginal colonization of SA and the antibiotic susceptibility of the isolates in pregnant women attending a maternity hospital in Cartagena, Colombia. Methods: Nasal and vaginal swabs were obtained from participants and subjected to microbiological and molecular assays. A post discharge follow-up was performed for up to four weeks. Results: From 100 pregnant women enrolled in the study, 34 were colonized with SA; 29 only in the nares, threeonly in the vagina, and two at both sites. Colonization of pregnant women with SA was more common in the nares than in the vagina or at both sites [29/34 (85.3%) vs 3/34 (8.8%) and 2/34 (5.9%); p

Dissertação de Mestrado Integrado em Medicina Veterinária Staphylococci can express a wide array of virulence traits which may strongly influence the infection prognosis, often being a multidrug resistant animal pathogen, responsible for conditions such as abscesses, dermatitis or mastitis. In this study we aimed to perform a phenotypic and genotypic characterization of virulence traits of methicillin-resistant staphylococci (MRS) isolates of animal origin. A total of 253 staphylococci were evaluated for methicillin resistance. This collection comprises isolates from bovine clinical and subclinical mastitis and other clinical isolates from dogs, cats, horses and goats. Identification had been previously performed by biochemical tests and confirmed to genus or species level by PCR and isolates clonality was evaluated by PFGE. Methicillin resistance screening was performed by the oxacillin disc diffusion according to CLSI guidelines and confirmed using a MRSA modified medium and by PCR amplification of the mecA gene. The occurrence of the new mecA homolog, LGA251 was evaluated by PCR in mecA negative MRS isolates. Virulence traits of MRS such as coagulase, haemolysins, DNase, gelatinase and lipase were phenotypically evaluated. Biofilm production was also phenotypically evaluated and by PCR of icaA, icaD and bap genes. Quorum-sensing system agr was determined by PCR. Antimicrobial susceptibility to 18 compounds was evaluated according to CLSI guidelines. The presence of bacteriophages and the production of bacteriocins were determined by plaque assays. A total of 26 isolates were identified as MRS: S. epidermidis (n=16), S. aureus (n=4) and Staphylococcus spp. (n=6). From the 26 MRS detected, 19.2% were coagulase-positive, 73.1% produced haemolysins, 26.9% produced DNase, 96.2% were gelatinase-positive, 69.2% were lipase-positive and 38.5% of the isolates were able to express biofilm in vitro. IcaA and icaD genes were both present in 38.5% of the isolates, but none was bap-positive. Only 11.5% of the isolates were typeable for agr. Antimicrobial co-resistance ranged from 0% (Chloramphenicol, Vancomycin) to 92.3% (Nalidixic acid). Bacteriophages were present in 34.6% of the isolates and none produced bacteriocins. The high frequencies of virulence traits present combined with the high antimicrobial coresistance profiles observed, suggest that these isolates may represent a serious problem with major public health implications, re-enforcing the importance of the wide concept of “One Health”. RESUMO - Caracterização fenotípica e genotípica de factores de virulência de staphylococci de origem animal - As bactérias pertencentes ao género Staphylococcus podem expressar uma ampla gama de factores de virulência que influenciam o prognóstico de uma infecção. Podem ser agentes patogénicos multirresistentes em animais, responsáveis por doenças como abcessos, dermatites ou mastites. Neste estudo realizou-se uma caracterização fenotípica e genotípica dos factores de virulência de staphylococci resistentes à meticilina (MRS) de origem animal. Um total de 253 staphylococci foram avaliados para resistência à meticilina. Esta colecção inclui isolados de mastites clínicas e subclínicas bovinas e isolados clínicos de cães, gatos, cavalos e cabras. A identificação dos isolados tinha sido previamente realizada por testes bioquímicos e foi confirmada ao género ou espécie por PCR e a clonalidade dos isolados foi avaliada por PFGE. A resistência à meticilina foi avaliada pelo método de difusão em disco com oxacilina de acordo com as normas do CLSI e confirmada pelo meio MRSA modificado e por PCR do gene mecA. A ocorrência do novo homólogo mecA, LGA251 foi avaliada por PCR em MRS mecA negativos. A presença de factores de virulência como coagulase, hemolisinas, DNase, gelatinase e lipase foi avaliada fenotipicamente. A produção de biofilme foi também avaliada fenotipicamente e por PCR dos genes icaA, icaD and bap. O sistema de quorum-sensing agr foi avaliado por PCR. A susceptibilidade a 18 antimicrobianos foi determinada de acordo com as normas do CLSI. A presença de bacteriófagos e a produção de bacteriocinas foram avaliadas por ensaios em placa. Um total de 26 isolados foram identificados como MRS: S. epidermidis (n=16), S. aureus (n=4) e Staphylococcus spp. (n=6). Dos 26 MRS, 19,2% eram coagulase-positivos, 73,1% produziram hemólise, 26,9% produziram DNase, 96,2% foram gelatinase-positivos, 69,2% foram lipase-positivos e 38,5% dos isolados foram capazes de expressar biofilme in vitro. Os genes icaA e icaD estão presentes em 38,5% dos isolados, mas nenhum foi bap-positivo. Apenas 11,5% dos isolados foram tipificados para com o sistema agr. A co-resistência a agentes antimicrobianos variou de 0% (cloranfenicol e vancomicina) até 92,3% (ácido nalidíxico). Os bacteriófagos estavam presentes em 34,6% dos isolados e nenhum produziu bacteriocinas. As elevadas frequências de factores de virulência combinadas com os elevados perfis de coresistência a antimicrobianos sugerem que estes isolados podem representar um problema sério para saúde pública, reforçando a importância do conceito de "Uma só saúde".

Objective Determining the prevalence of nasal carriage of S. aureus, both sensitive to methicillin and resistant to it, in preschool children and evaluating the presence of Panton-Valentine leukocidin genes in the isolates. Methods This was a cross-sectional study in which cultures from anterior nares were obtained from healthy preschool children. Isolates were identified as S. aureus based on morphological and biochemical tests. Antibiotic susceptibility profiles were determined by the disk diffusion method. All the isolates were further analyzed by multiplex PCR to determine the presence of mecA and PVL genes; methicillin-resistant isolates were also SCCmec typed by multiplex PCR. Results Overall S. aureus nasal colonization prevalence was 38.5 % and 4.8 % for methicillin-resistant strains. All the methicillin-resistant isolates carried the genes for PVL; two isolates possessed the SCCmec type IV, two were SCCmec type I and one was SCCmec type II. Conclusion This study revealed high PVL-positive, methicillin-resistant S. aureus colonization prevalence in healthy preschool children from Cartagena, which may play a key role in the epidemiology of community-associated infection by methicillin-resistant S. aureus in healthy children from this particular geographical area. Objetivo Determinar la prevalencia de colonización nasal de S. aureus, tanto sensible como resistente a meticilina, en niños preescolares y evaluar la presencia de los genes de la leucocidina Panton-Valentine en estos aislamientos. Métodos Estudio de corte transversal en el que se realizaron cultivos de flora nasal de niños preescolares. Los aislamientos fueron identificados como S. aureus con base en su morfología y pruebas bioquímicas. La susceptibilidad a antibióticos se determinó por el método de difusión en disco. Todos los aislamientos fueron analizados por PCR múltiple para determinar la presencia de los genes mecA y PVL, y para la tipificación del casete cromosómico SCCmec de los aislamientos resistentes a meticilina. Resultados La colonización nasal por S. aureus fue 38,5 %, y la de cepas meticilino-resistentes fue 4,8 %. Todos los aislamientos SARM portaban los genes para PVL, dos portaban el elemento SCCmec tipo IV, dos fueron tipo I y uno fue tipo II. Conclusión Encontramos una alta prevalencia de colonización por cepas meticilino-resistentes, PVL-positivos en la población estudiada, lo que podría jugar un papel clave en la epidemiología de las infecciones por S.aureus meticilino-resistente en esta área geográfica.

Staphylococcus aureus resistente à meticilina (MRSA), outrora isolado especificamente em ambientes hospitalares, vem sendo identificado como causador de infecções cutâneas em pacientes da comunidade. Neste artigo, é relatado um caso do sul do Brasil com furunculose por CA-MRSA. O microrganismo isolado foi submetido a exames de PCR para o gene mecA e para o gene que codifica a leucocidina de Panton-Valentine. Esses exames permitiram a identificação genotípica do CA-MRSA. Methicillin-resistant Staphylococcus aureus (MRSA), particularly isolated at hospital setting, has been identified in cutaneous infections of community patients. This paper reports a case of furunculosis from the southern Brazil. Dermatologists must be attentive to this emergent etiological diagnosis. The isolated microorganism was subjected to PCR for gene mecA and to PCR for the gene that encodes the leukocidin of Panton-Valentine. These exams enabled genotypic identification of CA-MRSA.

Methicillin-resistant Staphylococcus aureus (MRSA), particularly isolated at hospital setting, has been identified in cutaneous infections of community patients. This paper reports a case of furunculosis from the southern Brazil. Dermatologists must be attentive to this emergent etiological diagnosis. The isolated microorganism was subjected to PCR for gene mecA and to PCR for the gene that encodes the leukocidin of Panton-Valentine. These exams enabled genotypic identification of CA-MRSA.

PURPOSE: To evaluate different methods of oxacillin susceptibility testing of ocular isolates, considering polymerase chain reaction (PCR) as the 'gold standard', and to compare the in vitro susceptibility to oxacillin with that of other antimicrobials used in ophthalmologic practice. METHODS: The Vitek gram-positive identification card was used to identify ocular coagulase negative Staphylococcus species. The presence of the mecA gene was determined by the polymerase chain reaction assay with a combination of two primer sets (mecA and 16S rRNA) in a single region. Results were analyzed and compared with other oxacillin susceptibility methods: PBP2a detection by rapid slide latex agglutination test (SLA); oxacillin E-test; the Vitek automated gram-positive susceptibility card (GPS-105); the oxacillin salt agar screening test (OSAS) at a concentration of 6.0, 1.0 and 0.75 µg oxacillin per ml and the cefoxitin disk diffusion test (CDD). Automated susceptibility was also determined to other antimicrobial agents (fluoroquinolones, penicillin G, amoxicillin-ampicillin, cefazolin, ampicillin-sulbactam, erythromycin, clindamycin, gentamicin, tetracycline, trimethoprim-sulfamethoxazole, vancomycin and rifampin. RESULTS: Of the 69 CoNS isolates tested, 71% were mecA-positive and 29% mecA-negative. All methods tested had a statistically significant agreement with polymerase chain reaction. There was a tendency of positive polymerase chain reaction predomination among the S. epidermidis isolates in comparison to non-epidermidis isolates, although this was not statistically significant (78.3% vs. 56.5%; chi2= 2.54; P= 0.11). The oxacillin salt agar screening test (0.75 µg oxacillin/ml) showed the best performance, with 100% sensitivity and negative predictive value; 95% specificity and 98% positive predictive value. Using the E-test, the mecA-positive isolates were statistically significantly more resistant to ciprofloxacin, ofloxacin, gatifloxacin and moxifloxacin (P= 0.002; P= 0.008; P= 0.002 and P= 0.003, respectively). There was a statistically significant higher proportion of resistance of the coagulase negative Staphylococcus mecA-positives for: penicillin G, amoxicillin-ampicillin, cefazolin, ampicillin-sulbactam, erythromycin, clindamycin, gentamicin and tetracycline (P