[In vitro activity of ceftaroline against Spanish isolates of Staphylococcus aureus: a multicenter study]. / Estudio multicéntrico sobre la actividad in vitro de ceftarolina frente a Staphylococcus aureus aislados en España.

INTRODUCTION: Ceftaroline fosamil is a new-generation antimicrobial agent of cephalosporins subgroup. It is the first commercially available beta-lactam antibiotic that exhibits activity against methicillin-resistant Staphylococcus aureus (MRSA). The aim of this study is to determine the in vitro Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values of ceftaroline against S.aureus strains (including MRSA). MATERIAL AND METHODS: A multicenter study involving four hospitals representative of the Spanish geography was performed. MIC and MBC values against both the methicillin-resistant and sensitive strains of S.aureus (MRSA and methicillin-sensitive S.aureus [MSSA]) were determined using a broth microdilution method. RESULTS: A total of 266 S.aureus strains were analyzed (95 MRSA and 171 MSSA). Ceftaroline bacterial sensitivity showed a mean MIC of 0.227 µg/ml (SD=0.146; range, 0.06 to 1 µg/ml). All MIC values of the 266 strains tested belonged to the sensitive category (value ≤ 1 µg/ml). Intermediate or resistant strains were not detected. MIC50 and MIC90 values for MRSA were 0.25 and 0.5 µg/ml, respectively (range=0.125-1 µg/ml). MSSA strains showed MIC50 and MIC90 values of 0.125 and 0.25 µg/ml, respectively (range=0.125-0.5 µg/ml). MBC50 and MBC90 values for MRSA were 0.5 and 1 µg/ml, respectively (range=0.125-1 µg/ml). MSSA strains showed MBC50 and MBC90 values of 0.25 and 0.25 µg/ml, respectively (range=0.125-0.5 µg/ml). CONCLUSION: Ceftaroline shows excellent in vitro activity against S.aureus, including MRSA strains. Therefore, this antibiotic may be a promising alternative for the treatment of infections caused by this bacterium.

Th1 and Th17 hypercytokinemia as early host response signature in severe pandemic influenza.

INTRODUCTION: Human host immune response following infection with the new variant of A/H1N1 pandemic influenza virus (nvH1N1) is poorly understood. We utilize here systemic cytokine and antibody levels in evaluating differences in early immune response in both mild and severe patients infected with nvH1N1. METHODS: We profiled 29 cytokines and chemokines and evaluated the haemagglutination inhibition activity as quantitative and qualitative measurements of host immune responses in serum obtained during the first five days after symptoms onset, in two cohorts of nvH1N1 infected patients. Severe patients required hospitalization (n = 20), due to respiratory insufficiency (10 of them were admitted to the intensive care unit), while mild patients had exclusively flu-like symptoms (n = 15). A group of healthy donors was included as control (n = 15). Differences in levels of mediators between groups were assessed by using the non parametric U-Mann Whitney test. Association between variables was determined by calculating the Spearman correlation coefficient. Viral load was performed in serum by using real-time PCR targeting the neuraminidase gene. RESULTS: Increased levels of innate-immunity mediators (IP-10, MCP-1, MIP-1beta), and the absence of anti-nvH1N1 antibodies, characterized the early response to nvH1N1 infection in both hospitalized and mild patients. High systemic levels of type-II interferon (IFN-gamma) and also of a group of mediators involved in the development of T-helper 17 (IL-8, IL-9, IL-17, IL-6) and T-helper 1 (TNF-alpha, IL-15, IL-12p70) responses were exclusively found in hospitalized patients. IL-15, IL-12p70, IL-6 constituted a hallmark of critical illness in our study. A significant inverse association was found between IL-6, IL-8 and PaO2 in critical patients. CONCLUSIONS: While infection with the nvH1N1 induces a typical innate response in both mild and severe patients, severe disease with respiratory involvement is characterized by early secretion of Th17 and Th1 cytokines usually associated with cell mediated immunity but also commonly linked to the pathogenesis of autoimmune/inflammatory diseases. The exact role of Th1 and Th17 mediators in the evolution of nvH1N1 mild and severe disease merits further investigation as to the detrimental or beneficial role these cytokines play in severe illness.

[Influenza surveillance by molecular diagnosis]. / Vigilancia de la gripe mediante diagnóstico molecular.

INTRODUCTION: Our objective was to evaluate the application of molecular techniques in the surveillance of influenza, and to describe clinical and epidemiological characteristics of cases diagnosed in 2007-2008 and 2008-2009 seasons. METHODS: We analyzed 183 pharyngeal swabs from the same number of patients referred to the virology laboratory of the Sentinel Physician Network of Castilla y Leon, the study of influenza viruses by shell-vial technique and RT-PCR capable of detecting multiple Simultaneously, influenza virus A, B, C, respiratory syncytial virus A, B and adenovirus. RESULTS: Using cell culture were isolated 17 influenza A viruses and 19 influenza B viruses (19.7% of total). By multiple RT-PCR, was detected 49 influenza A virus, 29 influenza B virus, an influenza virus C, 3 syncytial virus type A and other B and 6 adenoviruses (44.3% of total). All influenza viruses isolated in cell culture was detected by RT-PCR. RT-PCR by 5 co-infections were detected, which represented a 6.25% of co-infections on the whole of positive samples. The average age of patients was 29 years (SD = 21.07). The proportion of women and men accounted for 43.7% and 56.3% respectively. The number of cases diagnosed in relation to age follows a pattern of negative linear correlation. CONCLUSIONS: RT-PCR is revealed as an useful tool for epidemiological surveillance of influenza, allowing also to detect viral subtypes along with other viruses involved in respiratory infections.

[Viral gastroenteritis. Application of a protocol for astrovirus detection in childhood gastroenteritis]. / Gastroenteritis de etiología vírica. Aplicación de un protocolo para el diagnóstico de astrovirus en casos de gastroenteritis infantil.

BACKGROUND AND OBJECTIVE: Diagnosis of viral gastroenteritis is an important subject in clinical virology which is mainly determined by the availability of reagents in laboratories, such as in the case of astrovirus. The aim of this study was to estimate the increase in the diagnostic performance achieved after the incorporation of astrovirus search in the diagnosis protocol of acute viral gastroenteritis. We also analyzed the trend of infections in other more commonly searched virus, such as rotavirus and enteric adenovirus. MATERIAL AND METHODS: Retrospective study during 20 years that included 12,980 stool samples processed for gastroenteritis virus diagnosis. Since 1997 an enzyme immunoassay for astrovirus has been applied to those samples that are negative for rotavirus and adenovirus. The study was divided in two periods (1986-1996 and 1997-2005, without and with astrovirus diagnosis) and the percentage of patients diagnosed in each period was compared. The trend of positive results as well as the percentage of positive results over all patients studied was modelled using the least squares method. RESULTS: The percentages of positive patients for rotavirus, adenovirus and astrovirus were 10.3%, 2.3% and 6.0% respectively, and there were uncommon co-infections by rotavirus and adenovirus (0.2%). The protocol applied to the astrovirus diagnosis increased the diagnosis rate up to 16.8% of the studied cases. Significant statistical differences were observed between the 2 study periods. A quadratic growth was observed in the results of positive diagnosis of viral gastroenteritis during the study period. CONCLUSIONS: The search of astrovirus in gastroenteritis cases by a selective protocol increased the diagnostic performance of gastrointestinal virus by 6%. In view of these results, it would be useful to implement astrovirus diagnosis in faeces with liquid or semi-liquid consistency when rotavirus and adenovirus detection is negative.

Comparación entre cinco técnicas de PCR para el diagnóstico del SARS-CoV-2 / Comparison between five PCR techniques for the diagnosis of SARS-CoV-2

Introducción. Desde que aparecieron los primeros casosde SARS-CoV-2 son numerosas las técnicas que se han desarrollado para el diagnóstico o seguimiento de la infección,tanto técnicas directas como serológicas. La elección de unabuena herramienta diagnóstica es fundamental para el controlepidemiológico. El objetivo ha sido comparar cinco técnicascomercializadas de RT-PCR a tiempo real, en sensibilidad, especificidad y concordancia para la detección del SARS-CoV-2.Material y métodos. Se compararon cinco kits comerciales de RT-PCR para la detección del SARS-CoV-2. Se tomaronocho muestras positivas conocidas que se sometieron a sietediluciones o concentraciones diferentes y otras 135 muestrasnegativas para determinar valores de sensibilidad, especificidad y concordancia.Resultados. La sensibilidad, especificidad, valor predictivopositivo (VPP) y valor predictivo negativo (VPN) para las técnicas de Palex, Roche y GeneXpert respecto a Seegene fueronidénticas, correspondientes a 98,21%, 100%, 100% y 99,26%respectivamente. Para Becton Dickinson la sensibilidad fue del89,28%, la especificidad del 100%, el VPP del 100% y el VPNdel 95,74%. La concordancia mediante el índice Kappa paraPalex, Roche y GeneXpert fue del 0,9892, mientras que la concordancia para Becton Dickinson fue con un índice Kappa de0,9215.Conclusión. Todos los kits de RT-PCR comerciales presentaron elevadas sensibilidades y especificidades, así como VPP,VPN y concordancia. (AU)

Introduction. Since the first cases of SARS-CoV-2appeared, there have been numerous techniques that havebeen developed for the diagnosis or monitoring of infection, both direct and serological techniques. Choosing agood diagnostic tool is essential for epidemiological control. The objective was to compare five commercializedRT-PCR techniques in real time, in sensitivity, specificityand agreement for the detection of SARS-CoV-2.Material and methods. Five commercial RT-PCR kitsfor the detection of SARS-CoV-2 were compared. Eightknown positive samples were taken and subjected to seven different dilutions or concentrations, and another 135negative samples were used to determine sensitivity, specificity, and agreement values.Results. The sensitivity, specificity, positive predictivevalue (PPV) and negative predictive value (NPV) for thePalex, Roche and GeneXpert techniques with respect toSeegene were identical, corresponding to 98.21%, 100%,100% and 99.26% respectively. For Becton Dickinson thesensitivity was 89.28%, the specificity of 100%, the PPVof 100% and the NPV of 95.74%. The agreement using theKappa index for Palex, Roche and GeneXpert was 0.9892,while the agreement for Becton Dickinson was with aKappa index of 0.9215.Conclusion. All commercial RT-PCR kits had highsensitivities and specificities, as well as PPV, NPV, andconcordance. (AU)

In vitro activity of ceftaroline against mecC-positive MRSA isolates.

Ceftaroline is a new cephalosporin with activity against methicillin-resistant Staphylococcus aureus (MRSA). A collection of 17 clinical and veterinary mecC-positive MRSA isolates was tested to evaluate the in vitro efficacy of ceftaroline against recently emerged mecC-MRSA isolates. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of ceftaroline for the 17 isolates were determined by broth microdilution using the methodology and interpretive criteria of the Clinical and Laboratory Standards Institute (CLSI). Additional susceptibility tests were performed using ceftaroline M.I.C.Evaluator (M.I.C.E.™) strips. All isolates showed susceptibility according to CLSI breakpoints, with MICs of ceftaroline ranging from 0.125mg/L to 0.25mg/L. MBCs were identical or up to a twofold dilution step higher. In conclusion, all tested isolates, from various sources and belonging to several clonal complexes (CCs), but predominantly to CC130, were found to be susceptible to ceftaroline. Ceftaroline could thus be an option for the treatment of mecC-MRSA infections.

Queratitis fúngica por Colletotrichum gloeosporioides: A propósito de un caso / Fungal keratitis caused by Colletotrichum gloeosporioides: A case report

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Blefaritis crónica grave. Evolución tórpida de varios años / Severe chronic blepharitis. Torpid evolution of several years

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Queratitis por Nocardia farcinica en paciente inmunocompetente. Descripción del primer caso en España / Keratitis by Nocardia farcinica in immunocompetent patient. Description of the first case in Spain

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Evaluación de un nuevo dispositivo para la recogida de la muestra, transporte y detección del estreptococo del grupo B en mujeres embarazadas / Evaluation of a new device for sample collection, transport and detection of Group B Streptococcus in pregnant women

Se ha diseñado un dispositivo de nueva invención que combina recogida, transporte, cultivo y detección del estreptococo del grupo B (EGB) sin necesidad de procesamiento ni manipulaciones intermedias, de manera que simplifique todo el proceso. El objetivo ha sido evaluar el rendimiento y utilidad de dicho dispositivo en la detección del EGB en mujeres embarazadas. Se comparó el nuevo prototipo en paralelo con la siembra directa de las muestras vagino-rectales en el medio sólido Granada en placas tradicionales. Mediante la siembra directa se detectaron 124 muestras positivas de 600 (20,6%). Mediante el nuevo dispositivo se detectaron las mismas que en siembra directa y además 10 adicionales 134/600 (22,3%). La utilización del nuevo dispositivo podría ser considerada en la práctica clínica asistencial de rutina para el cribado del EGB mediante previo acuerdo de comercialización (AU)

We have designed a new device that combines sample collection, transportation, culture and detection of Group B Streptococcus (GBS), requiring no additional processing in the clinical laboratory. The objective was to evaluate the performance of this device for GBS detection in pregnant women. The new prototype was compared to direct plating of vaginal-rectal swabs onto Granada solid media plates. Direct plating method detected 124 positive samples out of 600 (20.6%) whereas the new device detected 10 additional positive samples (134/600, 22.3%). This new device (patent-protected) could be considered for routine GBS screening (AU)

Desarrollo de una PCR para la detección y cuantificación de la parasitación por Demodex folliculorum en biopsias de neoplasias cutáneas del área periocular / Development of a PCR for the detection and quantification of parasitism by Demodex folliculorum infestation in biopsies of skin neoplasms periocular area

Objetivo. Estandarizar la cuantificación relativa por masa de tejido de la parasitación por Demodex folliculorum procedente de biopsias neoplásicas cutáneas de la zona periocular mediante técnicas de amplificación molecular con el objetivo de poder estudiar la posible relación de la aparición de carcinoma basocelular palpebral con la presencia y densidad del ácaro en trabajos posteriores. Material y métodos. Se desarrolló una PCR cuantitativa a tiempo real con sondas TaqMan. La PCR se probó en una serie piloto de 46 muestras reales de biopsias de carcinoma basocelular de tipo nodular. Resultados. La sensibilidad se situó con un límite de detección de entre 1 y 10 copias/μl. El 50% (23/46) de las biopsias fueron positivas a D. folliculorum. La especificidad fue del 100% confirmado mediante secuenciación. Conclusión. La técnica muestra buenos resultados de sensibilidad y especificidad que la pueden hacer útil como herramienta para estudios causa-efecto de D. folliculorum y basalioma (AU)

Objective. To standardize the relative quantification by mass of tissue parasitism by Demodex folliculorum infestation from neoplastic skin biopsies periocular using molecular amplification to study the possible relationship of the appearance of eyelid basal cell carcinoma with the presence and density of the mite in later works. Methods. A quantitative PCR was developed real-time probes TaqMan. PCR was tested in a pilot 46 actual biopsy samples nodular basal cell carcinoma series. Results. The sensitivity was placed with a detection limit of between 1 and 10 copies / μl. 50% (23/46) of the biopsies were positive for D. folliculorum. The specificity was 100% confirmed by sequencing. Conclusion. The technique shows good results for sensitivity and specificity that can make it useful as a tool for studies of cause and effect D. folliculorum and basal cell carcinoma (AU)

Optimización en la recuperación del estreptococo del grupo B en el cribado de mujeres embarazadas / Enhanced recovery of group B Streptoccocus in pregnant women screening

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