RESUMO
Vancomycin is the cornerstone in treating methicillin-resistant Staphylococcus aureus (MRSA) infections. However, therapeutic failures can occur when MRSA strains with decreased susceptibility to glycopeptides (DSG) are involved. The aim of this study was to detect and characterize DSG in MRSA recovered from children with invasive diseases at a reference pediatric hospital between 2009 and 2019. Fifty-two MRSA strains were screened using agar plates with vancomycin 3 and 4 mg/L (BHI-3 and BHI-4); the VITEK2 system; and standard and macro E-tests. Suspicious hVISA were studied by population analysis profiling-area under the curve (PAP-AUC), and wall thickness was analyzed by transmission electron microscopy. Neither VRSA nor VISA were detected in this set. As only three strains met the hVISA criteria, the PAP-AUC study included 12 additional MRSA strains that grew one colony on BHI-4 plates or showed minimum inhibitory concentrations of vancomycin and/or teicoplanin ≥ 1.5 mg/L. One strain was confirmed as hVISA by PAP-AUC. The wall thickness was greater than the vancomycin-susceptible control strain; it belonged to ST30 and carried SCCmec IV. As expected, a low frequency of hVISA was found (1.9%). The only hVISA confirmed by PAP-AUC was not detected by the screening methods, highlighting the challenge that its detection represents for microbiology laboratories.
RESUMO
Abstract The aim of this study was to characterize phenotypically and genotypically 27 mecApositive Staphylococcus aureus strains with oxacillin MICs of ≤2 g/ml by Vitek 2, isolated indifferent regions of Uruguay. Susceptibility to oxacillin and cefoxitin was studied by gradient dif-fusion, disk diffusion to cefoxitin, and Phoenix and MicroScan systems. PBP2a was determined.SCCmec typing was performed and the isolates were compared by PFGE. Twenty-six isolateswere susceptible to oxacillin; one strain was susceptible to cefoxitin by disk diffusion and 3strains by gradient diffusion. Phoenix and MicroScan panels detected methicillin resistance in25 and 27 strains, respectively. Twenty-six strains tested positive for PBP2a. Twenty-six strainscarried SCCmec V and 24 belonged to pulsotype A. One strain carried SCCmec IV and did notbelong to pulsotype A. Cefoxitin disk diffusion test and PBP2a detection correctly identified 26of these 27 strains as MRSA. PFGE results suggest the dissemination of a cluster of MRSA carryingSCCmec V.
Resumen El objetivo de este estudio fue caracterizar fenotípicamente y genotípicamente 27 cepas de Staphylococcus aureus positivas para mecA y con CIM de oxacilina <2 pg/ml según Vitek 2, obtenidas en diferentes regiones del país. La sensibilidad frente a la oxacilina y la cefoxitina se estudió por difusión en gradiente, por disco-difusión (cefoxitina) y por los sistemas Phoenix y MicroScan. Se analizó la portación de PBP2a, se realizó la tipificación de SCCmec y las cepas se compararon mediante PFGE. Resultaron sensibles a oxacilina por difusión en gradiente 26 cepas; una fue sensible a cefoxitina por disco-difusión y 3 lo fueron por difusión en gradiente. Los sistemas Phoenix y MicroScan detectaron resistencia a meticilina en 25 y 27 cepas, respectivamente. Asimismo, 26 cepas portaban PBP2a y 26 cepas mostraron presencia de SCCmec V, 24 correspondieron al pulsotipo A. Una portaba SCCmec IV y no perteneció al pulsotipo A. La prueba de disco-difusión con cefoxitina y la detección de PBP2a identificaron 26 de 27 cepas como MRSA. La PFGE sugiere la diseminación de un grupo MRSA con SCCmec V. © 2022 Asociación Argentina de Microbiología. Publicado por Elsevier Espana, S.L.U. Este es un artículo Open Access bajo la licencia CC BY-NC-ND (https://creativecommons.org/licenses/by-nc-nd/4.0/).
RESUMO
Introducción: Streptococcuspyogenes (EGA) es agente de enfermedad invasora (EI); su alta morbimortalidad exige vigilancia epidemiológica. Objetivo: Describir características clínicas y epidemiológicas de niños hospitalizados con EI por EGA en un centro de referencia de Uruguay del 1/1/2014 al 31/12/2020 incluyendo el estudio de los factores de virulencia encontrados en las cepas aisladas. Materiales y Métodos: Descriptivo y retrospectivo. Definición de caso: aislamiento de EGA. en sitios estériles. Variables: epidemiológicas, clínicas, laboratorio, tratamiento y evolución. Se tipificó por secuenciación del gen emm. Se obtuvieron perfiles cromosómicos por digestión del ADN con la enzima SmaI. Presencia de los genes que codifican SpeB, SpeA, SpeC y Ssa, y susceptibilidad a antimicrobianos. Resultados: Tasa de admisiones: 3,98/10.000. Se incluyeron 22 pacientes; infección osteoarticular (n = 11), infección pleuropulmonar (n = 6), absceso no cutáneo (n = 4) y aislamiento en sangre (n = 1). Media de edad: 44 meses; 8 fueron graves, siendo su media de edad menor (16 meses) Todas los casos con neumonías fueron graves y un paciente falleció. Se secuenciaron 12 cepas: 5 emm1 (4 emm 1.29 y 1 emm 1) y 1 de cada uno de los siguientes: emm 6.4, emm 81, emm12, emm28, emm 22, emm 87, emm 11. Todas eran SpeB+. Perfiles de toxinas: SpeA+SpeC-Ssa-(5), SpeA-SpeC+Ssa-(4) SpeA-SpeC-Ssa-(2) y SpeA-SpeC+Ssa+ (2). Conclusiones: Este estudio permite dar continuidad a un estudio previo. Se logró mayor tipificación de EGA. que puede contribuir a su conocimiento clínico molecular. No hubo registro de pacientes con diagnóstico de SST ni de fascitis necrosante, a diferencia de la serie anterior.
Background: Streptococcus pyogenes (GAS) is an agent of invasive disease (ID); its high morbidity and mortality requires epidemiological surveillance. Aim: To describe the clinical and epidemiological characteristics of children hospitalized with ID due to GAS in a reference center in Uruguay from January 1-2014 to December 31-2020, including a study of virulence factors. Methods: Descriptive and retrospective. Case definition: Isolation of GAS in sterile sites. Variables: epidemiological, clinical, laboratory, treatment and evolution. Strains were typified by sequencing of the emm gene. Chromosomal profiles were obtained by digestion of the DNA. with the Smal enzyme. Presence of SpeB, SpeA and SpeC genes and susceptibility to antibiotics were performed. Results: Admissions rate: 3.98/10,000. 22 patients were included; osteoarticular infection (n = 11), pleuropulmonary infection (n = 6), non-cutaneous abscess (n = 4) and blood isolation (n = 1). Mean age: 44 months; 8 cases were severe, their mean age was lower (16 months). All pneumonia cases were severe and one patient died. Twelve strains were sequenced: 5 emm1 (4 emm1.29 and 1 emm1) and 1 of each: emm6.4, emm81, emm 12, emm28, emm 22, emm 87, emm 11. All were SpeB+. Toxin profiles: SpeA+SpeC-Ssa-(5), SpeA-SpeC+Ssa-(4) SpeA-SpeC-Ssa-(2) and SpeA-SpeC+Ssa+(2). Conclusions: This study allows to give continuity to a previous study. Greater typing of GAS was achieved, which may contribute to its molecular clinical knowledge. There was no record of patients diagnosed with TSS or necrotizing fascitis, unlike the previous series.
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Resumen Los objetivos de este estudio fueron determinar el desempeño del panel BCID de FilmArray® y establecer el impacto de estos resultados en el tratamiento antimicrobiano de pacientes con bacteriemia en 11 hospitales de Latinoamérica. Se incluyeron 397 episodios de bacteriemia y se documentaron 551 microorganismos aislados de hemocultivos. La identificación microbiana fue correcta en el 91,4% (504/551) de los aislados y en el 98,6% (504/511) si se consideran solo los microorganismos incluidos en el panel BCID. La sensibilidad en la detección de los genes mecA, vanA/B y blaKPC fue del 100% y la especificidad fue del 97%, 100% y 99,6% respectivamente. La notificación temprana del resultado permitió cambios terapéuticos en 242 episodios (60,9%). El panel BCID es un método confiable y rápido para la detección de mecanismos críticos de resistencia y de los microorganismos más frecuentemente aislados de bacteriemias y permite la optimización temprana del tratamiento antimicrobiano.
Abstract The objectives of this study were to determine the performance of the BCID panel and to establish the impact of these results on the antimicrobial treatment of patients with bacteremia in 11 hospitals in Latin America. Three hundred and ninety-seven episodes of bacteremia were included and 551 microorganisms isolated from blood cultures were documented. Microbial identification was correct in 91.4% (504/551) of the isolates and in 98.6% (504/511) if only the microorganisms included in the BCID panel are considered. The sensitivity in the detection of the genes mecA, vanA/B and blaKPC was 100% and the specificity was 97%, 100% and 99.6% respectively. Early notification of the outcome allowed therapeutic changes in 242 episodes (60.9%). The BCID panel is a reliable and rapid method for the detection of critical resistance mechanisms and of the microorganisms most frequently isolated from bacteremia and it enables early optimisation of antimicrobial treatment.
Resumo Os objetivos deste estudo foram determinar o desempenho do painel BCID do FilmArray® e estabelecer o impacto desses resultados no tratamento antimicrobiano de pacientes com bacteremia em 11 hospitais da América Latina. Trezentos e noventa e sete episódios de bacteremia foram incluídos e 551 microrganismos isolados de hemoculturas foram documentados. A identificação microbiana foi correta em 91,4% (504/551) dos isolados e em 98,6% (504/511) considerando apenas os microrganismos incluídos no painel BCID. A sensibilidade na detecção dos genes mecA, vanA/B e blaKPC foi de 100% e a especificidade foi de 97%, 100% e 99,6% respectivamente. A notificação precoce do desfecho permitiu mudanças terapêuticas em 242 episódios (60,9%). O painel BCID é um método confiável e rápido para a detecção de mecanismos críticos de resistência e dos microrganismos mais frequentemente isolados da bacteremia e permite a otimização precoce do tratamento antimicrobiano.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Análise Custo-Eficiência , Bacteriemia/diagnóstico , Hemocultura/métodos , Anti-Infecciosos/farmacologiaRESUMO
The aim of this study was to characterize phenotypically and genotypically 27 mecA positive Staphylococcus aureus strains with oxacillin MICs of ≤2µg/ml by Vitek 2, isolated in different regions of Uruguay. Susceptibility to oxacillin and cefoxitin was studied by gradient diffusion, disk diffusion to cefoxitin, and Phoenix and MicroScan systems. PBP2a was determined. SCCmec typing was performed and the isolates were compared by PFGE. Twenty-six isolates were susceptible to oxacillin; one strain was susceptible to cefoxitin by disk diffusion and 3 strains by gradient diffusion. Phoenix and MicroScan panels detected methicillin resistance in 25 and 27 strains, respectively. Twenty-six strains tested positive for PBP2a. Twenty-six strains carried SCCmec V and 24 belonged to pulsotype A. One strain carried SCCmec IV and did not belong to pulsotype A. Cefoxitin disk diffusion test and PBP2a detection correctly identified 26 of these 27 strains as MRSA. PFGE results suggest the dissemination of a cluster of MRSA carrying SCCmec V.
Assuntos
Staphylococcus aureus Resistente à Meticilina , Infecções Estafilocócicas , Humanos , Oxacilina/farmacologia , Staphylococcus aureus , Cefoxitina/farmacologia , Uruguai , Antibacterianos/farmacologia , Proteínas de Bactérias/genética , Testes de Sensibilidade Microbiana , Staphylococcus aureus Resistente à Meticilina/genéticaRESUMO
BACKGROUND: Uruguay incorporated the conjugate vaccine against Haemophilus influenzae b (Hib) in 1994. In 2008, the vaccine was changed from one with natural conjugated capsular polysaccharide to one with a synthetic polysaccharide component. We describe the frequency and characteristics of invasive Hib infections in children hospitalized in a Pediatric Reference Hospital (PRH) between January 1st, 2000 and December 31st, 2017. METHODS: Sterile site Hib isolations from hospitalized children were included. Clinical and microbiological characteristics were analyzed. Favorable conditions for the infection were considered: incomplete immunization, immunodeficiencies and associated pathologies. Two periods are described: 1, prior to vaccine change (1/1 st/2000- 12/31/08) and 2, post-change (1/1 st/09- 12/31st/17). RESULTS: 45 children were hospitalized: 5 in the first period and 40 in the second. The hospitalization rate per 10,000 discharges was 0.41 (95% CI 0.05-0.77) and 4.2/10,000 (95% CI 2.89-5.48), respectively (p < 0.01). The diagnoses at discharge were: meningitis/ventriculitis (20), pneumonia (16), bacteremia (3), epiglottitis (1), arthritis (1), cellulitis (3) and obstruction of the upper airway (1). Four children presented comorbidities. Twenty seven received less than 3 doses of anti-Hib vaccination and 18 were properly vaccinated (2 were immunodeficient). The median hospitalization was 14 days, 18 children required intensive therapy. CONCLUSIONS: Observed change may be due to: incomplete primary series, inhomogeneous vaccine coverage and immunogenicity of the synthetic polysaccharide. To reduce this public health problem, epidemiological surveillance.
