Validación de la reacción de polimerasa en cadena en tiempo real como herramienta diagnóstica en la aspergilosis asociada a COVID-19 en pacientes hospitalizados en Clínica Dávila de Santiago, Chile, durante los años 2020 y 2021

Introducción: Desde el inicio de la pandemia por COVID-19 se han registrado casos de infecciones de aspergilosis pulmonar asociada a esta infección, la cual tiene características diferentes a la aspergilosis pulmonar clásica y, por lo tanto, han significado un desafío diagnóstico. Objetivo: Validar una reacción de polimerasa en cadena (RPC) en tiempo real (sigla en inglés RT-PCR) comercial, como herramienta diagnóstica alternativa a la técnica de galactomanano (GM) en el diagnóstico de aspergilosis pulmonar asociada a COVID-19 (sigla en inglés CAPA). Pacientes y Método: Se analizaron resultados de RT-PCR de Aspergillus spp y GM en lavado bronco-alveolar (LBA) de 72 pacientes, hospitalizados por COVID-19 de Clínica Dávila entre los años 2020 y 2021. De estos pacientes, 33 presentaron CAPA. Resultados: La RT-PCR de Aspergillus y GM presentaron una correlación positiva (r = 0,6351, valor p < 0,0001). La técnica de RT-PCR presentó una sensibilidad (S), especificidad (E), valor predictor positivo (VPP) y valor predictor negativo (VPN) de 100, 44, 66 y 100%, respectivamente, mientras que en GM fueron de 64, 89, 84 y 73%, respectivamente para LBA. Al utilizar ambas técnicas en combinación se obtuvo una S, E, VPP y VPN de 100, 82, 88 y 100%, respectivamente. Conclusión: Este estudio concluyó que usar una técnica de RT-PCR de Aspergillus y GM en conjunto en LBA mejoraron los parámetros de desempeño de ambas técnicas usadas de manera individual para diagnosticar CAPA. Se requieren más estudios para evaluar el desempeño de técnicas combinadas en otros tipos de aspergilosis.

Background: Since the beginning of the COVID-19 pandemic, there have been cases of pulmonary aspergillosis infections associated with this infection, which has different characteristics from classical pulmonary aspergillosis and therefore, have been diagnostic challenges. Aim: To validate a commercial real-time PCR (RT-PCR) method as an alternative diagnostic tool to the galactomannan (GM) technique in the diagnosis of COVID-19-associated pulmonary aspergillosis (CAPA). Methods: Results of RT-PCR of Aspergillus spp and GM in broncho-alveolar lavage (BAL) of 72 patients hospitalized for COVID-19 at Clínica Dávila between 2020 and 2021 were analyzed. Of these patients, 33 presented CAPA. Results: The RT-PCR for Aspergillus and GM showed a positive correlation (r = 0.6351, p-value < 0.0001). The RT-PCR for Aspergillus technique presented a sensitivity (S), specificity (S), positive predictive value (PPV) and negative predictive value (NPV) of 100, 44, 66 and 100% respectively, while the GM technique presented 64, 89, 84 and 73%, respectively for BAL. Using both techniques in combination a S, E, PPV and NPV of 100, 82, 88 and 100% were obtained respectively. Conclusion: This study concluded that using RT-PCR and GM techniques in combination in BAL improved the performance parameters of both techniques from those used individually to diagnose CAPA. Further studies are required to evaluate the performance of combined techniques in other aspergillosis focus.

Evaluación de la prueba Aspergillus Galactomannan Ag VircliaR Monotest como alternativa a kit Platelia ™ Aspergillus EIA / Evaluation of the Aspergillus Galactomannan ag VircliaR Monotest test as an alternative to Platelia™ Aspergillus EIA kit

INTRODUCCIÓN: La prueba Aspergillus galactomannan Ag Virclia® (GM-VClia) es una técnica de galactomanano monotest, auto-matizada, basada en inmunoensayo quimioluminiscente (CLIA). OBJETIVO: Evaluar el desempeño del test de GM-VClia en muestras de suero y lavado bronquioalveolar (LBA) procesadas previamente con el kit Platelia™ Aspergillus EIA (GM-Plat). MATERIALES Y MÉTODOS: Se estudiaron 56 muestras de suero y 40 de LBA, correspondientes a un total de 59 pacientes (algunos con determinación de galactomamano en ambas muestras) con enfermedades pulmonares, hematológicas, LES, Covid-19 y tumores, entre otros. Trece pacientes tuvieron aspergilosis invasora (1 probada y 12 probables). RESULTADOS: La correlación entre ambos métodos para suero y LBA fue r = 0,8861 p < 0,0001 y r = 0,6368 p < 0,001, respectivamente. Hubo una concordancia global de 67,7% (65/96), siendo de 85,7% (48/56) en sueros y 42,5,0% (14/49) en LBA. Al subir el punto de corte en LBA por GM-VClia la concordancia aumentó a 85,7%. CONCLUSIONES: Se observó una mayor correlación y concordancia en sueros que en LBA. El kit GM-VClia presentó una mayor sensibilidad y valor predictor negativo (VPN), que el kit GM-Plat. Las desventajas de GM-VClia, la constituyen la categoría "dudoso", que dificulta la interpretación y que, con los puntos de corte actuales en LBA, la correlación con GM-Plat es menor. Las ventajas son su mayor sensibilidad, facilidad de procesamiento y una mayor rapidez en los resultados.

BACKGROUND: The Aspergillus Galactomannan Ag Virclia® (GMVClia) test is a monotest and automated galactomannan technique based on chemiluminescent immunoassay (CLIA). AIM: To evaluate the performance of the GM-VClia test in serum and bronchioalveolar lavage (BAL) samples previously processed with the Platelia ™ Aspergillus EIA kit (GM-Plat). METHODS: 56 samples of serum 40 from BAL (some of them with galactomaman determination in both samples), from patients with pulmonary diseases, hematological diseases, SLE, Covid-19 and tumors, among others, were studied. Thirteen patients had invasive aspergillosis (1 proven and 12 probable). RESULTS: The correlation between both methods for serum and BAL was r = 0.8861 p < 0.0001 and r = 0.6368 p < 0.001, respectively. There was a global concordance of 67.7% (65/96), being 85.7% (48/56) in sera and 42.5.0% (14/49) in BAL. By raising the cut-off point in LBA by GM-VClia, the agreement increased to 85.7%. CONCLUSION: A greater correlation and concordance was observed in sera than in BAL. The GM-VClia kit had a higher sensitivity and NPV than the GM-Plat kit. The disadvantages of GM-VClia are the "doubtful" category, which makes interpretation difficult and that with the current cut-off points in LBA the correlation with GM-Plat is lower. The advantages are its greater sensitivity, ease of processing and faster results.

Adultos con influenza, evolución clínica, costos y grupos relacionados por el diagnóstico, resultados de 4 años. Clínica Dávila. Santiago de Chile / Adults patients with influenza, clinical evolution, cost and diagnosis related groups, four years results. Clinica Dávila, Santiago, Chile

Resumen Introducción: En 2009 la Influenza A H1N1pdm09 provocó en Chile 12.258 casos y 155 muertes. Objetivo: Analizar en adultos egresados de Clínica Dávila con influenza, en 2009, 2010, 2012 y 2014, soporte ventilatorio, costo de hospitalización, Grupos Relacionados por el Diagnóstico (GRD) y letalidad. Material y Método: Estudio descriptivo retrospectivo usando la ficha médica electrónica. Resultados: Egresaron 115.673 adultos, 338 (0,29%) con diagnóstico de Influenza, edad 56,5 ± 22 años, 59% mujeres, letalidad 4%. Hubo 3 grupos, Grupo 1: sin ningún soporte ventilatorio, 295 pacientes, edad 63 ± 20, estadía 6,6 ± 6,9 días, costo promedio de hospitalización $2.885.261, mediana peso GRD 0,41 (p25 = 0,38 y p75 = 0,62), letalidad 1,01% (3 pacientes). Grupo 2: Ventilación mecánica no invasiva (VMNI), 23 casos, edad 77,1 ± 13, letalidad 22% (5 casos), estadía 16,8 ± 12,4, costo $9.245.242, GRD 0,79 (p25 = 0,62 y p75=1,03). Grupo 3: Intubación y ventilación mecánica invasiva (VMI), 20 pacientes, edad 56,4 ± 15, estadía 36,9 ± 41,4, costo $38.681.099, GRD 5,86 (p25 = 5,82 y p75 = 5,86) y letalidad 30% (6 pacientes). Los GRD grupo VMI versus grupo VMNI y ningún soporte fueron diferentes (p < 0,0001 y p < 0,0001 respectivamente). La letalidad por influenza el 2014 fue de 8,5%, mientras que en los años 2012, 2010 y 2009 fue 1,5%, 3% y 2,5% respectivamente. La mediana de edad el año 2009 fue 37,5 años, menor que la de los otros años (p < 0,0001). Conclusiones: En 2009 los pacientes fueron más jóvenes, la necesidad de soporte ventilatorio provocó un peso GRD, estadía, costo y letalidad mayores que aquellos que no lo requirieron.

Introduction: In 2009 Influenza A H1N1pdm09 caused in Chile 12,258 cases and 155 deaths. Objective: To analyze ventilatory support, cost of hospitalization, Diagnosis Related Groups (DRG) and lethality in adults patients with influenza discharged from our institution, during 2009, 2010, 2012 and 2014. Patients and Method: Retrospective descriptive study using electronic medical records. Results: 115,673 adults were discharged, 338 (0.29%) with diagnosis of Influenza, age 56.5 ± 22 yr.o., 59% women, lethality 4%. There were 3 groups, Group 1: without any ventilatory support, 295 patients, age 63 ± 20, stay 6.6 ± 6.9 days, average cost of hospitalization 2,885,261 clp, medium weight DRG 0.41 (p25 = 0.38) andp75 = 0.62), lethality 1.01% (3 patients). Group 2: Non-invasive mechanical ventilation (NIMV), 23 cases, age 77.1 ± 13, lethality 22% (5 cases), stay 16.8 ± 12.4, cost 9,245,242 clp, DRG 0.79 (p25 = 0.62 and p75 = 1.03). Group 3: Intubation and invasive mechanical ventilation (IMV), 20 patients, age 56.4 ± 15, stay 36.9 ± 41.4, cost 38.681.099 clp, DRG 5.86 (p25 = 5.82 and p75 = 5,86) and lethality 30% (6 patients). The DRG group VMI versus group VMNI and no support were different (p < 0.0001 and p < 0.0001 respectively). The lethality for influenza in 2014 was 8.5%, while in 2012, 2010 and 2009 it was 1.5%, 3% and 2.5% respectively. The median age in 2009 was 37.5 yr.o significantly minor, than the other years (p < 0.0001). Conclusions: In 2009 the patients were younger, the need for ventilatory support led to a higher DRG weight, stay, cost and lethality than those who did not require it.

Interacción Candida albicans-Hospedero: un proceso complejo en el que la inmunidad innata juega un importante papel / Candida albicans-host interaction: a complex process in which the innate immunity play an important role

Candida albicans es una levadura comensal, que bajo ciertas circunstancia puede convertirse en patógeno. La capacidad de cambiar se forma constituye un factor de virulencia, permitiéndole a la levadura invadir y diseminarse. El sistema inmune innato es capaz de reconocer las diferentes morfologías de C. albicans activando receptores (PRRs) que reconocen PAMPs o patrones moleculares conservados. Las células inmunes más importantes son los macrófagos y los neutrófilos que desencadenan una respuesta efectora a través de la fagocitosis y la activación de estrés oxidativo contra C. albicans. Las células dendríticas, por su parte expresan la mayoría de los PRRs involucrados en el reconocimiento de C. albicans activando la secreción de citoquinas hacia una respuesta tipo TH1 (inducida por INF tipo1, IL-12, INFγ, ), Treg (inducida por TGFβ, IL-10) y TH17 (inducida por IL-23, IL6). Las células epiteliales no sólo constituyen una barrera física frente a la infección por C. albicans, sino son fundamentales en el reconocimiento primario de este microorganismo. Mediante una respuesta bifásica estas células activan diferencialmente, vías de transducción de señales que determinan que no se active una respuesta de citoquinas frente a la presencia de blastoconidios (forma comensal) y que se active frente a presencia de hifas (forma invasora). Por su parte, C. albicans es capaz de desarrollar mecanismos evasivos de la respuesta inmune. Esta compleja interacción y hongo-hospedero determina si el hospedero será capaz de eliminar a este microorganismo o si éste finalmente invadirá expresando su virulencia.

Candida albicans is a comensal microorganism that under certain circumstances is able to transform into a pathogen. This ability to switch constitute a virulence factor that C. albicans uses to invade and spread. The innate immune system recognize the different forms of C. albicans activating receptors (PRRs) that recognize PAMPs or conserved molecular patterns. The most important immune cells are macrophages and neutrophils that generate an effector response through phagocytosis and oxidative burst against C. albicans. Dendritic cells express the most of PRRs involved in the recognition of C. albicans activating the cytokines synthesis forward to a TH1 (induced by INF tipo1, IL-12, INFγ), Treg (induced by TGFβ, IL-10) y TH17 (induced by IL- 23, IL6) immune response. Epithelial cells not only constitute a physical barrier against C. albicans, but they are crucial in the first recognition of this microorganism. Through a biphasic response these cells differentially activate pathways that determine a no cytokine response when blastoconidia (commensal form) is present and an inflammatory response in presence of hyphae (pathogenic form). C. albicans develops immune evasive mechanism. This complex host-fungi interaction determines if the host will eradicate the infection or if this microorganism will invade the host, expressing it virulence.

Recomendaciones para el análisis de datos acumulados de susceptibilidad antimicrobiana en instituciones de salud / Recommendations for the analysis of cumulated data in antimicrobial susceptibility in health institutions

Due to the great variability in antimicrobial resistance patterns, local reports of cumulative antimicrobial susceptibility data are necessary in every health center. The purpose is to guide clinical decisions and the early detection of patterns that allow preventive measures to avoid dissemination of resistant strains. The main objective of this guide is to provide recommendations for the analysis of antimicrobial susceptibility data and elaboration of a local report. Recommendations provided in this guide are based on the Clinical and Laboratory Standards Institute (CLSI) document "Analysis and Presentation of Cumulative Antimicrobial Susceptibility Test Data" (3). Key aspects related to information gathering and data processing, analysis and presentation are described.

Considerando la gran variabilidad en la distribución de la resistencia microbiana, es una necesidad que cada centro de salud genere reportes locales de datos acumulados de susceptibilidad, con el propósito de guiar las decisiones clínicas y detectar tendencias que permitan establecer medidas de prevención para evitar la diseminación de cepas resistentes. Esta guía tiene como objetivo entregar recomendaciones para el análisis de susceptibilidad antimicrobiana y aportar datos útiles para la elaboración del informe local. Las recomendaciones que contenidas em este documento están basadas en el documento "Analysis and Presentation of Cumulative Antimicrobial Susceptibility Test Data de Clinical and Laboratory Standards Institute (CLSI) (3). Se describen aspectos claves relacionados con los requerimientos de la información, el procesamiento de los datos, el análisis y presentación de éstos.

Efecto antifúngico de quitosán de alto peso molecular en cepas de Candida sp aisladas de muestras clínicas / Antifungal effect of high molecular weight chitosan on Candida spp isolated from clinical samples

Chitosan is a D-glucosamine polysaccharide derived from chitin that displays an antimicrobial activity against bacteria and fungi. Objective: to evaluate the antifungal effect of high molecular weight chitosan (HMWC) in clinical strains of Candida spp. Methodology: the susceptibility of forty strains of Candida spp. to HMWC was studied (16 C albicans, 11 C glabrata, 5 C. tropicalis, 5 C krusei, 2 C parapsilosis and 2 C.famatd) by broth microdilution at pH 7.0 and pH 4.0. Results: of 40 strains, only 2 were inhibited at pH 7.0 and corresponded to ATCC control strains (C. krusei 6258 and C parapsilosis 22019). On the other hand, 37/40 strains (92.5 percent) were inhibited by concentrations lower than 1.25 mg/mL of HMWC at pH 4.0. Conclusion: these results show that HMWC, presents activity against clinical Candida spp. strains, including C glabrata, and that this activity is present at acid pH (4.0). This compound could potentially be used in vulvovaginal candidiasis since it occurs at pH 4.0-4.5.

Distribución y susceptibilidad a fluconazol de levaduras del género Candida aisladas en pacientes hospitalizados y ambulatorios / Species distribution and fluconazole susceptibility of yeasts of genus Candida isolated from hospitalized and ambulatory patients

Most surveillance studies have included invasive candidiasis from hospitalized patients. However, no national study has evaluated the species distribution and susceptibility to fluconazole of Candida species isolated from hospitalized and ambulatory patients. Atotal of 166 strains were collected consecutively during a 6 month period. Strains were isolated from vaginal fluid (73.5 percent), uriñe (7.8 percent), lower respiratory tract samples (7.8 percent), blood cultures (4.2 percent), sterile fluids (2.4 percent) and wounds (1.8 percent). Most of the isolates were obtained from ambulatory patients (71.1 percent). The species found were Candida albicans (78.9 percent), C glabrata (8.4 percent), C. tropicalis (6.0 percento), C.famata (1.8 percent), C krusei (1.8 percent), C parapsilosis (1.8 percent) and C. sake (1.2 percent). Fluconazole susceptibility was: 92.3 percento for C. albicans, 85.7 percent for C glabrata (most strains being dose-dependent susceptible), 100 percent) for C parapsilosis and 80 percent) for C tropicalis. Only susceptible strains were isolated from hospitalized children, whereas more resistant strains were isolated from ambulatory adults, mainly from vaginal fluid. In order to identify probable reservoirs of less susceptible strains such as C. glabrata, it would be necessary to include ambulatory isolates in future surveillance studies.

La mayoría de los estudios de vigilancia de levaduras del género Candida se realizan en candidiasis invasora en pacientes hospitalizados; sin embargo, no existen estudios que evalúen simultáneamente la distribución global de especies y susceptibilidad a fluconazol de cepas de Candida aisladas de pacientes hospitalizados y ambulatorios. Se analizaron 166 cepas de Candida, recolectadas consecutivamente en un período de 6 meses, las cuales fueron aisladas de flujo vaginal (73,5 por ciento), orina (7,8 por ciento), muestras respiratorias bajas (7,8 por ciento), hemocultivos (4,2 por ciento), líquidos estériles (2,4 por ciento) y heridas (1,8 por ciento). Un 71,1 por ciento provenía de pacientes ambulatorios. La especies aisladas fueron Candida albicans (78,9 por ciento), C glabrata (8,4 por ciento), C tropicalis (6,0 por ciento), Cfamata (1,8 por ciento), C. krusei (1,8 por ciento), C parapsilosis (1,8 por ciento) y C. sake (1,2 por ciento). Los porcentajes de sensibilidad a fluconazol fueron: 92,3 por cientoo para C. albicans, 85,7 por cientoo para C glabrata (siendo la mayoría sensible dosis dependiente), 100 por ciento) para C parapsilosis y 80 por cientoo para C tropicalis. En los niños, todos hospitalizados, se aislaron sólo cepas sensibles, mientras que en adultos ambulatorios se aislaron más cepas resistentes, fundamentalmente de flujo vaginal. Considerar cepas ambulatorias en futuros estudios de vigilancia permitirían dar cuenta de probables reservónos de cepas más resistentes, como C. glabrata.

Infección rinosinusal por Scedosporium apiospermum en un hospedero inmunocompetente / Scedosporium apiospermum rhinosinusal infection in an immunocompetent host

Scedosporium species can cause colonization, superficial and deep localized infection or systemic disease, espe-cially in irnmunocompromised hosts. We report a case of localized infection due to Scedosporium apiospermum in a 47 year oíd woman, with previous nasal surgery. She consulted for recurrent mucopurulent post-nasal discharge not responding to antibiotics. Computed tomography showed opacification of right maxillary sinus. Surgery was performed to removed abnormal tissue from sinus; biopsy revealed chronic sinusitis with aggregate of tightly packed hyphae suggestive of filamentous fungi. The microbiology fungal culture reported Scedosporium apiospermum.

Las infecciones por Scedosporium sp pueden traducirse en colonización, infecciones localizadas superficiales y profundas, o enfermedad diseminada. Presentamos un caso clínico de infección rinosinusal por Scedosporium apiospermum en una paciente de 47 años, con antecedente de cirugía por cuerpo extraño en la fosa nasal derecha. Consultó por descarga posterior muco-purulenta y recurrente, sin respuesta a tratamiento antibacteriano. Las imágenes de cavidades paranasales mostraron opacidad del seno maxilar derecho. Se realizó cirugía de remoción de contenido sinusal cuyo estudio histológico reveló sinusitis crónica erosiva, colonias de hongos con morfología sugerente de hongo filamentoso y desarrollo de S. apiospermum en el cultivo.

Estudio multicéntrico de la vigilancia de la susceptibilidad in vitro a tigeciclina en Santiago de Chile / Multicenter study on the monitoring of in vitro susceptibility to tigeeyeline in Santiago, Chile

The objective of this multicenter study was to determine tigeeyeline susceptibility rates, measured by agar diffusion, in nine hospitals in Santiago and to compare these rates with other antimicrobials. Each center studied 20 strains per month. All intermedíate and fully resistant strains as well as 10 percent of susceptibile strains were also studied by the broth microdilution method. Overall, 2301 strains were studied displaying the foliowing susceptibility rates for tigeeyeline: 100 percent for Streptococcus sp, Enterococcus sp, and E. coli respectively, 99.8 percent for Staphylococcus sp, 93 percent for Klebsiella and 80 percent for Acinetobacter baumarmii. For Proteus, Providencia and Morganella the susceptibility rates were 4 percent. For cefotaxime-resistant Klebsiella and imipenem-resistant A. baumarmii susceptibility rates were 95 percent and 80 percent respectively. The agar diffusion and broth dilution method were 100 percent concordant for tigeeyeline susceptible strains but only 27 percent for resistant or intermedíate strains represented mostly by Acinetobacter baumannii. The majority of these strains (57/59) proved to be susceptible after retesting. The great majority (96,6 percent) of strains tested from nine Chilean hospitals proved to be susceptible to tigeeyeline with exception for Proteus, Providencia and Morganella (66 percent resistance). Using the agar diffusion method for measuring tigeeyeline susceptibility to A. baumannii may be misleading.

Para conocer la susceptibilidad a tigeciclina por difusión en agar en nueve hospitales de Santiago y comparar la susceptibilidad con otros antimicrobianos, se diseñó este estudio multicéntrico. Cada centro estudió 20 cepas mensualmente. Las intermedias, resistentes y 10 por cientoo de las susceptibles se re-testearon y estudiaron por microdilución en caldo. Se incluyeron 2.304 cepas. Fueron susceptibles a tigeciclina Strep-tococcus sp (100 por cientoo), Enterococcus sp (100 por ciento), E. coli (100 por cientoo), Staphylococcus sp (99,8 por ciento), Klebsiella pneumoniae (93 por ciento) y Acinetobacter baumannii (80 por ciento). En Proteus, Providencia y Morganella la susceptibilidad fue 4 por cientoo. Klebsiella resistente a cefotaxima y Acinetobacter resistente a imipenem, 95 por cientoo y 80 por cientoo fueron susceptibles a tigeciclina, respectivamente. La concordancia en cepas susceptibles y en las enviadas como resistentes o intermedias (A. baumannii) fue 100 por cientoo y 27 por cientoo respectivamente. El re-testeo confirmó que la mayoría eran susceptibles. Los patrones de susceptibilidad bacteriana muestran muy buena actividad in vitro a tigeciclina. La resistencia in vitro de A. baumannii por difusión en agar debe interpretarse con precaución.

Actualización en pruebas de susceptibilidad antifúngica / An update on antifungal susceptibility testing

Due to increasing of invasive fungal infections and emergeney of antifungal drugs resistant fungi, standardized methods of antifungal susceptibility testing (AST) have been developed. The Clinical Laboratory Standards Instutute (CLSI) and the European for Committee Antimicrobial Susceptibility Testing (EUCAST) have guidelines for susceptibility of yeasts by broth microdilution (M27-A2 and E. Dis. 7.1 documents, respectively). Both are equivalent, although they present methodological and interpretative breakpoints differences. In addition, the CLSI have the M38-A (for filamentous fungi) and M44-A (disk diffusion) documents, whereas EUCAST is developing a document for Aspergillus spp. Furthermore, commercial methods are available that display good correlation with the methods of reference such as E-test®, Sensititre® and Vitek2®. The interpretation of the results must be careful because the determination of the minimum inhibitory concentration (CIM) is difficult for fungi, there are host factors involved and not always there is a correlation between MIC and clinical outeome. Due to these methods are laborious and require trained personnel, to ask for AST to a reference laboratory is recommendable.

Debido al aumento en las infecciones fúngicas invasores y a la emergencia de hongos resistentes a los antifúngicos, ha sido necesario desarrollar métodos estandarizados de susceptibilidad antifúngica. El Clinical Laboratory Standards Instutute (CLSI) y el European Committee for Antimicrobial Susceptibility Testing (EUCAST) han elaborado guías para susceptibilidad de levaduras por microdilución en caldo (documentos M27-A2 y E. Dis. 7.1, respectivamente). Ambos son equivalentes, aunque presentan diferencias metodológicas y en sus puntos de corte. El CLSI ha desarrollado los documentos M38-A (hongos filamentosos) y M44-A (difusión en disco), mientras que EUCAST trabaja en un documento para Aspergillus sp. Por otra parte, existen métodos comerciales que presentan buena correlación con los métodos de referencia como E-test®, Sensititre® y Vitek2®. La interpretación de los resultados debe ser cuidadosa pues la determinación de la concentración inhibitoria mínima (CIM) es muy dificultosa para hongos, hay factores del hospedero involucrados y no siempre hay una correlación entre la CIM y la respuesta a tratamiento. Como estas técnicas, en general, son laboriosas y requieren de personal entrenado, es recomendable derivar los estudios de susceptibilidad a un laboratorio de referencia.

Bacteriemia primaria por Listeria monocytogenes en paciente con cirrosis hepática: Caso clínico / Primary Listeria monocytogenes infection in a cirrhotic woman: Report of one case

L. monocytogenes infections are infrequent. Sepsis in pregnant women and newborns and central nervous system infections in the elderly are the most common clinical manifestations. We report a 61 years old woman with diabetes Mellitus and a Child B hepatic cirrhosis, admitted for persistent fever. Blood cultures were positive for Listeria monocytogenes. Cerebrospinal fluid was normal and sterile. She was treated with ampicillin and amikacin with a good response. Control blood cultures were negative. She was discharged 14 days after in good conditions.

Epidemiología clínica y molecular de las infecciones por Shigella spp en niños de la Región Metropolitana durante el verano 2004-2005 / Shigella spp Infections In children living In the Metropolitan RegionChile, during summer of 2004-2005

Background: Shigella spp is a frequent cause of diarrhea in children. Antimicrobials decrease the duration of diarrhea and pathogen excretion. However, the increasing resistance limits their therapeutic value. Aim: To study Shigella serotype distribution in the Metropolitan Region in Chile, and its relationship with severity of disease, antimicrobial resistance pattern and clonality. Material and methods: During summer 2004-2005, stool samples from children with diarrhea were collected in Cary Blair transpon medium and cultured. Shigella isolates were serotyped using monoclonal and polyclonal commercial antibodies. In vitro activity of ampicillin, amoxicillin/clavulanic acid, chloramphenicol, cotrimoxazol, nalidixic acid, ciprofloxacin, ceftriaxone and azythromycin was determined by minimal inhibitory concentration (MIC). Clonality was studied by pulsed-field gel electrophoresis (PFGE) using Xbal as restriction enzyme. Results: One hundred thirty nine Shigella strains were isolated (77 S sonnei and 62 S flexneri). S sonnei and S flexneri 2a serotypes were responsible for 95 percent of episodes. Children aged 2-4 years, showed a greater incidence of Shigella infections and 77 percent of episodes were treated on an ambulatory basis. High resistance levels were observed for ampicillin, cotrimoxazole, amoxicillin-clavulanic acid and chloramphenicol (67 percent, 60 percent, 56 percent and 45 percent, respectively). We found 11 resistance patterns and 61,2 percent of strains were multiresistant. There were multiple clones without a strict relationship with resistance patterns. Conclusions: Shigella infections in Metropolitan Region in Chile are associated to a restricted number of serotypes, representing a clonal expansion associated to different antimicrobial resistant patterns.

Suceptibilidad antifúngica de levaduras mediante etest©: comparación de 3 medios / Antifungal susceptibility testing by etest©: comparison of 3 media

Background: The increasing frequency of systemic fungal infections and the emergence of secondary resistance to antifungals in the lasts years, has stimulated the use of methods for antifungal susceptibility testing. Etest(r) is an easily performed quantitative method that has a good agreement with the broth microdilution reference method (NCCLS), if appropriate media are used. Aim: To compare the susceptibility to Amphotericin B (AmB) and Fluconazole (Flu) of 22 opportunistic yeast isolates (C albicans (7), C tropicalis (9), C parapsilosis (3) and Cryptococcus neoformans (3) by Etest ©., using three different media and to choose the best medium for each tested drug. The studied media were RPMI 1640, Casitone (Cas) and Sabouraud. Results: The interpretation of minimal inhibitory concentration (MIC) endpoints on Sabouraud was difficult for AmB. The American Type Culture Collection (ATCC) strains MICs were out of the acceptable range in this medium. RPMI and Cas were suitable media to test AmB and Flu, but best endpoints were obtained for AmB in RPMI and Flu in Cas. Conclusions: The use of appropriate media for each antifungal drug optimizes the MIC readings by Etest(r). AmB should be tested in RMPI and Flu in Cas. Sabouraud must not be used

Infecciones sistémicas por levaduras en un hospital general: correlación entre estudio de susceptibilidad in vitro y supervivencia de los pacientes al episodio de infección fúngica / Systemic yeast infections in a general hospital: correlation between antifungal susceptibility and patient survival

Background: Fungi are important causal agents of nosocomial infections, that usually have high mortality rates. Aim: To evaluate the species distribution and susceptibility patterns of deep yeast infections in a General Hospital and to correlate those results with patient survival. Material and methods: Twenty one strains (from five pediatric and 16 from adult patients) were studied. Antifungal Susceptibility Testing (AST) to Amphotericin B (Anfb), Fluorocytosine (5FC), Fluconazole (FZ) and Itraconazole (IZ) was performed according to the EUCAST document. Clinical data of patients was obtained and survival to the infection was recorded. Results: C. albicans was isolated in 11 samples (52 percent), C. parapsilosis in three samples (14 percent), C. glabrata in two samples (9 percent), C. tropicalis in one sample (5 percent) and C. neoformans in four samples (19 percent). Twenty three percent of fungi were recovered at the Surgical Intensive Care Unit. The MICs ranged between 0.25 and 0.5 µg/mL for Anfb; between 0.25 and 16 µg/ml for SFC, between 0.12 and 32 µg/mL for FZ and 0.015 and 0.5 µg/mL for IZ. No association between antifungal susceptibility and patient survival was observed. Conclusions: C. albicans continues to be the most frequently isolated yeast, however, non-albicans species are an emergent group causing nosocomial infections. Surgical procedures are the main source of fungal infections in this sample

Valores de referencia de inmunoglobulina A secretora (IgAs) en saliva de niño sano / Reference values of secretory immunoglobuline A in saliva of healthy children

Objetivo: describir la relación entre las concentraciones normales de IgAs y la albúmina salival en niños normales. Material y método: se midieron los concentraciones de IgAs, albúmina y razón 19AS/albúmina en saliva de 60 niños sanos de 0 a 9 años. Resultados: la concentración de IgAs no se distribuyó normalmente en la población estudiada y la distribución no se normalizó al corregir por albúmina. Los puntos de corte establecidos en base a los percentile, 2,5 y 97,5 en el conjunto de la muestra fueron 40,2 y 788 mg/L respectivamente, siendo el promedio de 239,03 m/L. No hubo diferencia estadísticamente significativa entre grupos por edad. Conclusiones: Aún no existe suficiente información sobre la utilidad clínica de la razón IgAs/albúmina, la cual debe evaluarse