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INTRODUCTION: Despite antifungal advancements, candidaemia still has a high mortality rate of up to 40%. The ECMM Candida III study in Europe investigated the changing epidemiology and outcomes of candidaemia for better understanding and management of these infections. METHODS: In this observational cohort study, participating hospitals enrolled the first ten consecutive adults with blood culture-proven candidemia. Collected data included patient demographics, risk factors, hospital stay duration (follow-up of 90 days), diagnostic procedures, causative Candida spp., management details, and outcome. Controls were included in a 1:1 fashion from the same hospitals. The matching process ensured similarity in age (10-year range), primary underlying disease, hospitalization in intensive care versus non-ICU ward, and major surgery within 2 weeks before candidemia between cases and controls. Overall and attributable mortality were described, and a survival probability for cases and controls was performed. RESULTS: One hundred seventy-one pairs consisting of patients with candidemia and matched controls from 28 institutions were included. In those with candidemia, overall mortality was 40.4%. Attributable mortality was 18.1% overall but differed between causative Candida species (7.7% for Candida albicans, 23.7% for Candida glabrata/Nakaseomyces glabratus, 7.7% for Candida parapsilosis and 63.6% for Candida tropicalis). Regarding risk factors, the presence of a central venous catheter, total parenteral nutrition and acute or chronic renal disease were significantly more common in cases versus controls. Duration of hospitalization, and especially that of ICU stay, was significantly longer in candidemia cases (20 (IQR 10-33) vs 15 days (IQR 7-28); p = 0.004). CONCLUSIONS: Although overall and attributable mortality in this subgroup analysis of matched case/control pairs remains high, the attributable mortality appears to have decreased in comparison to historical cohorts. This decrease may be driven by improved prognosis of Candida albicans and Candida parapsilosis candidemia; whereas candidemia due to other Candida spp. exhibits a much higher attributable mortality.
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Candida , Candidemia , Humanos , Candidemia/mortalidad , Candidemia/microbiología , Masculino , Femenino , Persona de Mediana Edad , Europa (Continente)/epidemiología , Anciano , Factores de Riesgo , Estudios de Cohortes , Candida/aislamiento & purificación , Candida/clasificación , Adulto , Anciano de 80 o más Años , Antifúngicos/uso terapéutico , Estudios de Casos y ControlesRESUMEN
BACKGROUND: To date, azoles represent the only viable option for oral treatment of invasive Candida infections, while rates of azole resistance among non-albicans Candida spp. continue to increase. The objective of this sub-analysis of the European multicenter observational cohort study Candida III was to describe demographical and clinical characteristics of the cohort requiring prolonged hospitalization solely to complete intravenous (iv) antifungal treatment (AF Tx). METHODS: Each participating hospital (number of eligible hospitals per country determined by population size) included the first ~ 10 blood culture proven adult candidemia cases occurring consecutively after July 1st, 2018, and treating physicians answered the question on whether hospital stay was prolonged only for completion of intravenous antifungal therapy. Descriptive analyses as well as binary logistic regression was used to assess for predictors of prolonged hospitalization solely to complete iv AF Tx. FINDINGS: Hospital stay was prolonged solely for the completion of iv AF Tx in 16% (100/621) of candidemia cases by a median of 16 days (IQR 8 - 28). In the multivariable model, initial echinocandin treatment was a positive predictor for prolonged hospitalization to complete iv AF Tx (aOR 2.87, 95% CI 1.55 - 5.32, p < 0.001), while (i) neutropenia, (ii) intensive care unit admission, (iii) catheter related candidemia, (iv) total parenteral nutrition, and (v) C. parapsilosis as causative pathogen were found to be negative predictors (aOR 0.22 - 0.45; p < 0.03). INTERPRETATION: Hospital stays were prolonged due to need of iv AF Tx in 16% of patients with candidemia. Those patients were more likely to receive echinocandins as initial treatment and were less severely ill and less likely infected with C. parapsilosis.
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Candida , Candidemia , Adulto , Humanos , Antifúngicos/uso terapéutico , Candidemia/microbiología , Tiempo de Internación , Equinocandinas/uso terapéutico , Estudios de Cohortes , Azoles/uso terapéutico , Candida parapsilosis , Factores de RiesgoRESUMEN
BACKGROUND: The European Confederation of Medical Mycology (ECMM) collected data on epidemiology, risk factors, treatment, and outcomes of patients with culture-proven candidaemia across Europe to assess how adherence to guideline recommendations is associated with outcomes. METHODS: In this observational cohort study, 64 participating hospitals located in 20 European countries, with the number of eligible hospitals per country determined by population size, included the first ten consecutive adults with culture-proven candidaemia after July 1, 2018, and entered data into the ECMM Candida Registry (FungiScope CandiReg). We assessed ECMM Quality of Clinical Candidaemia Management (EQUAL Candida) scores reflecting adherence to recommendations of the European Society of Clinical Microbiology and Infectious Diseases and the Infectious Diseases Society of America guidelines. FINDINGS: 632 patients with candidaemia were included from 64 institutions. Overall 90-day mortality was 43% (265/617), and increasing age, intensive care unit admission, point increases in the Charlson comorbidity index score, and Candida tropicalis as causative pathogen were independent baseline predictors of mortality in Cox regression analysis. EQUAL Candida score remained an independent predictor of mortality in the multivariable Cox regression analyses after adjusting for the baseline predictors, even after restricting the analysis to patients who survived for more than 7 days after diagnosis (adjusted hazard ratio 1·08 [95% CI 1·04-1·11; p<0·0001] in patients with a central venous catheter and 1·09 [1·05-1·13; p<0·0001] in those without one, per one score point decrease). Median duration of hospital stay was 15 days (IQR 4-30) after diagnosis of candidaemia and was extended specifically for completion of parenteral therapy in 100 (16%) of 621 patients. Initial echinocandin treatment was associated with lower overall mortality and longer duration of hospital stay among survivors than treatment with other antifungals. INTERPRETATION: Although overall mortality in patients with candidaemia was high, our study indicates that adherence to clinical guideline recommendations, reflected by higher EQUAL Candida scores, might increase survival. New antifungals, with similar activity as current echinocandins but with longer half-lives or oral bioavailability, are needed to reduce duration of hospital stay. FUNDING: Scynexis.
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Candida , Candidemia , Adulto , Humanos , Antifúngicos/uso terapéutico , Adhesión a Directriz , Candidemia/tratamiento farmacológico , Candidemia/epidemiología , Candidemia/microbiología , Europa (Continente)/epidemiología , Estudios de CohortesRESUMEN
We analyzed the species distribution and susceptibility patterns of 433 strains of Aspergillus spp. isolated from respiratory samples of 419 in-patients included in multicenter prospective study (FUNGAE-IFI) between July 2014 and October 2015. Identification was carried out by conventional methods at each participating center and by molecular sequencing of a portion of the ß-tubulin gene at one of the centers. In vitro susceptibility was evaluated by broth microdilution methods and using the E-test (for cryptic species). Species identified included 249 A. fumigatus sensu stricto, 60 A. terreus sensu stricto, 47 A. flavus sensu stricto, 44 A. tubingensis, 18 A. niger sensu stricto , five A. nidulans sensu stricto, three A. tamarii, two A. calidoustus, two A. carneus, one A. acuelatus, one A. carbonarius, and one A. sydowii. Cryptic species were found in 12.5% of isolates (n = 54). The frequency of non-wild-type isolates for amphotericin B was 3.4% (n = 15) of the isolates tested and for azoles 3% (n = 10). None of the Aspergillus spp. were non-wild type to echinocandins. Of the 54 cryptic species only two strains were non-wild-type strains by microdilution method (3.7%) (two A. tubingensis, one to amphotericin B and another one to voriconazole) and by E-test method five strains of A. tubingensis showed high minimal inhibitory concentration (MIC) to amphotericin B (11.4%) and five to azoles (12.1%), one A. calidoustus strain showed high MICs for three azoles (50%), A. carneus to itraconazole (100%) and A. sydowii to amphotericin B and itraconazole (100%). These results provide relevant information on susceptibility patterns, frequency, and epidemiology of species involved in respiratory tract samples and of the incidence of recently described cryptic species.
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Antifúngicos/farmacología , Aspergilosis/epidemiología , Aspergilosis/microbiología , Aspergillus/clasificación , Aspergillus/aislamiento & purificación , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/microbiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Aspergillus/efectos de los fármacos , Niño , Preescolar , Femenino , Humanos , Incidencia , Lactante , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Estudios Prospectivos , Análisis de Secuencia de ADN , Encuestas y Cuestionarios , Tubulina (Proteína)/genética , Adulto JovenRESUMEN
INTRODUCTION: The genus Aspergillus contains more than 300 species, which are divided into closely related groups called sections. Molecular studies have revealed numerous cryptic species within different sections of this genus, which have different profiles of antifungal susceptibility and lack diagnostic morphological features. However, there are few studies on the prevalence and in vitro antifungal susceptibility of the cryptic species of this genus. The aim of this study was to investigate the distribution of Aspergillus spp. among clinical samples, and to study their in vitro susceptibility to different antifungal drugs. METHOD: Over a period of 2-years (2014-2015), a total of 379 strains of the genus Aspergillus were isolated. Most of the isolates were classified as respiratory colonizations; no cases of invasive aspergillosis were found. The strains were identified by MALDI-TOF mass spectrometry, and susceptibility testing was performed by the EUCAST reference procedure. RESULTS: Twenty species belonging to 8 sections were identified, being A. fumigatus the most prevalent (44.1%). The prevalence of cryptic species was 15.3%, with a clear predominance of A. tubingensis. Among the tested antifungal drugs, amphotericin B was the less active in vitro, followed by triazole drugs and echinocandins. The cryptic species had minimun inhibitory concentrations (MICs) higher than the corresponding type species. CONCLUSIONS: Accurate identification of the genus Aspergillus at the species level and in vitro antifungal susceptibility testing are necessary because, as it has been shown, some species of this genus may show resistance profiles against available antifungal drugs.
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Antifúngicos/farmacología , Aspergillus/efectos de los fármacos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Aspergillus/clasificación , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Adulto JovenRESUMEN
OBJECTIVE: Patients in PICUs frequently present hypochloremic metabolic alkalosis secondary to loop diuretic treatment, especially those undergoing cardiac surgery. This study evaluates the effectiveness of acetazolamide therapy for metabolic alkalosis in PICU patients. DESIGN: Retrospective, observational study. SETTING: A tertiary care children's hospital PICU. PATIENTS: Children receiving at least a 2-day course of enteral acetazolamide. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Demographic variables, diuretic treatment and doses of acetazolamide, urine output, serum electrolytes, urea and creatinine, acid-base excess, pH, and use of mechanical ventilation during treatment were collected. Patients were studied according to their pathology (postoperative cardiac surgery, decompensated heart failure, or respiratory disease). A total of 78 episodes in 58 patients were identified: 48 were carried out in cardiac postoperative patients, 22 in decompensated heart failure, and eight in respiratory patients. All patients received loop diuretics. A decrease in pH and PCO2 in the first 72 hours, a decrease in serum HCO3 (mean, 4.65 ± 4.83; p < 0.001), and an increase in anion gap values were observed. Urine output increased in cardiac postoperative patients (4.5 ± 2.2 vs 5.1 ± 2.0; p = 0.020), whereas diuretic treatment was reduced in cardiac patients. There was no significant difference in serum electrolytes, blood urea, creatinine, nor chloride after the administration of acetazolamide from baseline. Acetazolamide treatment was well tolerated in all patients. CONCLUSIONS: Acetazolamide decreases serum HCO3 and PCO2 in PICU cardiac patients with metabolic alkalosis secondary to diuretic therapy. Cardiac postoperative patients present a significant increase in urine output after acetazolamide treatment.
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Acetazolamida/uso terapéutico , Alcalosis/tratamiento farmacológico , Inhibidores de Anhidrasa Carbónica/uso terapéutico , Cuidados Críticos/métodos , Adolescente , Niño , Preescolar , Esquema de Medicación , Femenino , Humanos , Lactante , Recién Nacido , Unidades de Cuidado Intensivo Pediátrico , Modelos Lineales , Masculino , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Protozoan parasites of the genus Marteilia have been detected in marine bivalves and other invertebrates around the world, associated in some cases with mass mortalities. The present paper reports the characterization of the Marteilia sp. protozoan infecting the digestive gland of the Grooved Razor Shell clam (Solen marginatus) from Galicia (NW Spain), proposing a novel species in the genus: Marteilia octospora n. sp. Morphological and molecular techniques were used for the description of this parasite. Tissue imprints were essential in the study to confirm the presence of 8 spores per sporangium, a number never reported in other species from this genus. An ultrastructural study revealed that the size and number of dense granules, free in the mature sporangia, were quite different from granules in other Marteilia spp. Another morphological difference is the absence of a layer of concentric membranes found surrounding the mature spore in other species. In addition, concentric membranous structures observed in the different stages of the parasite have never been mentioned in other species of genus Marteilia. Moreover, molecular analysis of the rDNA intergenic spacer (IGS) and the internal transcribed spacer (ITS-1) showed differences with the sequences available for other Marteilia spp.
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Bivalvos/parasitología , Cercozoos/fisiología , Animales , Bivalvos/ultraestructura , Cercozoos/clasificación , Cercozoos/genética , Cercozoos/crecimiento & desarrollo , Microscopía Electrónica de Transmisión , Filogenia , ARN Ribosómico 18S/genética , ARN Ribosómico 28S/genética , España , Esporas Protozoarias/ultraestructuraRESUMEN
OBJECTIVE: We investigated the temporal pattern and predictive value of neutrophil gelatinase-associated lipocalin for early identification of acute kidney injury in children undergoing cardiac surgery. DESIGN: Prospective observational cohort study. SETTING: One PICU in a tertiary medical center in Madrid, Spain. PATIENTS: One hundred six children older than 15 days and younger than 16 years undergoing surgery for congenital cardiac lesions. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Urine samples were obtained before and at intervals after surgery. Acute kidney injury was defined according to pediatric Risk, Injury, Failure, Loss, and End-stage kidney disease criteria. The temporal pattern of both urine neutrophil gelatinase-associated lipocalin absolute concentration elevation and normalized to urine creatinine concentration was correlated with the development of acute kidney injury and other clinical outcomes. We evaluated the predictive ability of both urine neutrophil gelatinase-associated lipocalin and urine neutrophil gelatinase-associated lipocalin/creatinine by area under the curve, when added to a clinical predictive model. Data from 106 pediatric patients were analyzed. Acute kidney injury occurred in 42 patients (39.6%). Urine neutrophil gelatinase-associated lipocalin significantly increased in patients with acute kidney injury at 1, 3, and 15 hours postoperatively. Urine neutrophil gelatinase-associated lipocalin and urine neutrophil gelatinase-associated lipocalin/creatinine correlated with surgical variables and clinical outcomes. Acute kidney injury prediction improved when urine neutrophil gelatinase-associated lipocalin was added to a clinical model (area under the curve increased at 1 hr from 0.85 to 0.91 and at 3 hr to 0.92). Neither the urine neutrophil gelatinase-associated lipocalin nor the urine neutrophil gelatinase-associated lipocalin/creatinine values were significantly different between patients with prerenal and sustained acute kidney injury. CONCLUSIONS: Urine neutrophil gelatinase-associated lipocalin is a predictive biomarker for acute kidney injury after pediatric cardiac surgery, and it may permit earlier intervention that improves outcome of acute kidney injury. Urine neutrophil gelatinase-associated lipocalin normalized to urine creatinine improves the prediction of acute kidney injury severity but offers no advantage in acute kidney injury diagnosis.
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Lesión Renal Aguda/orina , Proteínas de Fase Aguda/orina , Biomarcadores/orina , Procedimientos Quirúrgicos Cardíacos/efectos adversos , Lipocalinas/orina , Proteínas Proto-Oncogénicas/orina , Lesión Renal Aguda/diagnóstico , Lesión Renal Aguda/etiología , Adolescente , Niño , Preescolar , Estudios de Cohortes , Femenino , Humanos , Lactante , Recién Nacido , Unidades de Cuidado Intensivo Pediátrico , Lipocalina 2 , Masculino , Periodo Posoperatorio , Estudios Prospectivos , EspañaRESUMEN
The aim of the present study was to carry out a survey of parasites and other conditions affecting pod razor clam populations, Ensis siliqua, in two beds from Galicia (NW Spain). In Galicia, the production of E. siliqua has increased in recent years due to the development of specific plans for its exploitation, however few and quite recent pathological studies have been carried out in this species. The results of this study showed the presence of different protozoa as the more prevalent group, especially Nematopsis sp. gregarines, unidentified branchial protozoa, renal coccidia and Trichodina sp. ciliates. Larval stages of trematodes and neoplastic disorders were also observed with lower prevalences. Furthermore, an ultrastructural analysis of two types of unidentified basophilic inclusions, both found in the digestive gland, revealed the presence of icosahedral viral particles and prokaryotic organisms, respectively. None of the parasites detected in E. siliqua from this study was notifiable to the World Organisation for Animal Health (OIE) and the majority of the symbionts and conditions observed in their tissues did not cause host damage. Nevertheless, parasites like bucephalid digenean sporocysts, viral inclusions, prokaryotic infections, disseminated neoplasm or germinoma detected in some samples could cause moderate or severe damage to the host depending on the intensity of infection.
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Bivalvos/parasitología , Mariscos/parasitología , Animales , España , SimbiosisRESUMEN
The aim of this study was to determine whether renal unresponsiveness to aldosterone associated with hyperkalemia is present in infants with acute pyelonephritis in the absence of significant urinary tract anomalies and to describe the clinical characteristics of patients presenting an inadequate renal response to hyperkalemia. The patient cohort comprised 113 infants with acute pyelonephritis (APN), based on the criteria of a temperature >38°C and significant bacteriuria. Serum and urine electrolytes, creatinine, osmolality, and renal tubular function tests were performed at diagnosis. The findings were compared to those present in 75 children who had fever without significant bacteriuria. Hyperkalemia (>5.5 mmol/L) was observed in infants with an APN diagnosis, who exhibited a lower transtubular potassium concentration gradient (TTKG) and a higher fractional sodium excretion. We defined inadequate renal response to hyperkalemia as the combination of hyperkalemia and TTKG below the normal range established for the age of the subject. Infants presenting an inadequate response to hyperkalemia were younger and associated more frequently with an APN diagnosis. This alteration could be explained by the renal interstitial inflammation present in acute pyelonephritis and the immaturity of the renal tubular responsiveness to aldosterone due to infancy in the absence of urinary tract infection or obstruction.
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Pielonefritis/metabolismo , Desequilibrio Hidroelectrolítico/etiología , Enfermedad Aguda , Femenino , Humanos , Lactante , Túbulos Renales/metabolismo , Masculino , Potasio/metabolismo , Estudios Prospectivos , Seudohipoaldosteronismo/etiologíaRESUMEN
BACKGROUND: There is little clinical information about community-onset bloodstream infections (COBSIs) caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESBLEC). We investigated the prevalence and risk factors for COBSI due to ESBLEC, and described their clinical features and the impact of COBSI caused by ESBLEC on 14-day mortality. METHODS: Risk factors were assessed using a multicenter case-control-control study. Influence of ESBL production on mortality was studied in all patients with COBSI due to E. coli. Isolates and ESBLs were microbiologically characterized. Statistical analysis was performed using multivariate logistic regression. Thirteen tertiary care Spanish hospitals participated in the study. RESULTS: We included 95 case patients with COBSI due to ESBLEC, which accounted for 7.3% of all COBSI due to E. coli. The ESBL in 83 of these (87%) belonged to the CTX-M family of ESBL, and most were clonally unrelated. Comparison with both control groups disclosed association with health care (odds ratio [OR], 2.1; 95% confidence interval [CI], 1.2-3.8), urinary catheter use (OR, 3.1; 95% CI, 1.5-6.5), and previous antimicrobial use (OR, 2.7; 95% CI, 1.5-4.9) as independent risk factors for COBSI due to ESBLEC. Mortality among patients with COBSI due to ESBLEC was lower among patients who received empirical therapy with beta-lactam/beta-lactam inhibitor combinations or carbapenems (8%-12%) than among those receiving cephalosporins or fluoroquinolones (24% and 29%, respectively). Mortality among patients with COBSI due to E. coli was associated with inappropriate empirical therapy irrespective of ESBL production. CONCLUSIONS: ESBLEC is an important cause of COBSI due to E. coli. Clinicians should consider adequate empirical therapy with coverage of these pathogens for patients with risk factors.
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Bacteriemia/microbiología , Infecciones Comunitarias Adquiridas/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli/enzimología , beta-Lactamasas/biosíntesis , Adolescente , Adulto , Anciano , Antibacterianos/farmacología , Bacteriemia/epidemiología , Estudios de Casos y Controles , Infecciones Comunitarias Adquiridas/epidemiología , Escherichia coli/efectos de los fármacos , Escherichia coli/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Femenino , Humanos , Modelos Logísticos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Análisis Multivariante , Pronóstico , Factores de Riesgo , España/epidemiología , Resistencia betalactámicaRESUMEN
Symbionts and abnormal conditions of razor clam Ensis arcuatus were surveyed in three commercially important natural beds of Galician estuaries (NW Spain). Samples of 15-20 E. arcuatus were collected every 2 months from January 2003 until July 2004 and processed for histological examination. Prokaryote-like colonies, renal coccidians, gregarines, Trichodina sp. ciliates, haplosporidian-like plasmodia, turbelaria, trematode metacercariae, cestode-like larvae and basophilic inclusion bodies were observed in razor clam tissues without causing host damage. Bucephalid digenean sporocysts and germinoma were seen in some samples causing moderate or severe damage to the host depending on the intensity of infection and both could be a cause for concern if prevalence reached epizootic levels in Galician E. arcuatus populations. None of the parasites detected is OIE notifiable and, in general, the commercially exploited beds studied seem to be devoid of serious pathogens.
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Bivalvos/microbiología , Animales , Infecciones por Protozoos/epidemiología , España , Simbiosis , Infecciones por Trematodos/epidemiología , Infecciones por Trematodos/veterinariaRESUMEN
OBJECTIVES: The aim of this study was to determine the antifungal drug susceptibilities of Candida bloodstream isolates in Andalusia, obtained through a multicenter active laboratory-based surveillance between October 2005 and September 2006. METHODS: One hundred and ninety-seven Candida isolates were collected. The MICs of amphotericin B, fluconazole, itraconazole and voriconazole were established using the Sensititre YeastOne panel. The MICs of posaconazole and caspofungin were determined by Etest. RESULTS: C. albicans was the most frequently isolated species (49.2%), followed by C. parapsilosis (17.3%), C. tropicalis (15.2%), C. glabrata (13.7%) and C. krusei (3.6%). All strains were inhibited at MICs of =1 mg/L of amphotericin B and 98.5% of isolates were inhibited at MICs of < or = 1 mg/L of posaconazole. A total of 8 isolates (4.1%) were classified as resistant to fluconazole (MIC > or = 64 mg/L) and 7 (3.6%) were considered resistant to itraconazole (MIC > or = 1 mg/L). All the isolates were susceptible to voriconazole and caspofungin. CONCLUSION: In our study C. krusei and C. glabrata were identified in over 18% of cases of candidemia. Most clinical isolates of these species are resistant or susceptible-dose-dependent to fluconazole but susceptible to voriconazole and caspofungin. These agents must be used in the empiric treatment of candidemia rather than fluconazole.
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Antifúngicos/farmacología , Candida/efectos de los fármacos , Candidiasis/microbiología , Farmacorresistencia Fúngica , Fungemia/microbiología , Adulto , Candida/aislamiento & purificación , Candidiasis/epidemiología , Farmacorresistencia Fúngica Múltiple , Fungemia/epidemiología , Humanos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Vigilancia de la Población , España/epidemiología , Especificidad de la EspecieRESUMEN
INTRODUCTION: The incidence of infections has decreased in kidney transplant (KT) recipients owing to advances in the surgical techniques and clinical management of this population. Nevertheless, these complications continue to occur and the causes seem to be changing, in part because of the prophylactic strategies used. METHOD: Prospective, observational study investigating infections occurring during the first 2 years post-transplantation in KT recipients who underwent surgery between July 2003 and December 2005 at Hospital Universitario Virgen del Rocío. Univariate and multivariate regression analysis was performed to determine risk factors associated with the development of infection. RESULTS: The incidence of infection was 1.11 episodes per patient over 510+/-234 days. The most common infections were urinary tract infection (UTI) (46.6%), cytomegalovirus (CMV) infection (22.7%), and surgical site infection (8%). The causes were bacterial (50.4%), viral (45.9%), and fungal (3.6%) agents. The most frequent pathogens were CMV (36%), Escherichia coli (28%), extended-spectrum beta-lactamase (ESBL)-producers (26%), and coagulase-negative staphylococci (6.3%). Seventy-nine percent of infection episodes occurred in the 4 months following KT. One recipient died 30 days after the infection episode. In the infection group, patient and graft survival at the end of follow-up was 98% and 89%, respectively. CONCLUSIONS: The most frequent syndromes were UTI, CMV infection and surgical site infection. The infections were mainly produced by bacteria, in particular gram-negative rods, and there was a high rate of ESBL E. coli.
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Enfermedades Transmisibles/epidemiología , Trasplante de Riñón , Complicaciones Posoperatorias/epidemiología , Adulto , Anciano , Anciano de 80 o más Años , Profilaxis Antibiótica , Bacteriemia/epidemiología , Infecciones por Citomegalovirus/epidemiología , Femenino , Infecciones por Bacterias Gramnegativas/epidemiología , Infecciones por Bacterias Grampositivas/epidemiología , Hospitales Universitarios/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Micosis/epidemiología , Neumonía/epidemiología , España/epidemiología , Infección de la Herida Quirúrgica/epidemiología , Infecciones Urinarias/epidemiología , Virosis/epidemiología , Adulto JovenRESUMEN
The definite and rapid diagnosis of invasive aspergillosis is necessary because of the high mortality caused. The objective of this study was to evaluate a real-time PCR assay to detect Aspergillus spp. in clinical samples, based on the Light Cycler technology. Specificity was assessed by using DNA extracted from pathogenic and non-pathogenic bacteria/fungi from Spanish Collection including: two Aspergillus flavus, four Aspergillus fumigatus, two Aspergillus nidulans, two Aspergillus niger and two Aspergillus terreus isolates. The analytical sensitivity was evaluated with different inocula (10(1)-10(5) conidia ml(-1)), and serially diluted DNA of A. fumigatus. To assess clinical applicability, samples from patients at risk were analysed. Species identification was determined by analysing the melting curves. Reactions using genomic DNA from other species of different genera than Aspergillus were negative (specificity: 100%). Analytical sensitivity was 60 fg using DNA and 5-20 conidia using conidial suspensions. The linear range was from 60 to 6 x 10(7) fg. The Tm ranged from 67.34 to 70.7 degrees C for the different Aspergillus spp. studied. Nine hundred and forty-eight consecutive blood samples from 127 patients were processed. In total, 10 (1%) of 948 samples from blood samples were PCR-positive. The real-time PCR assay provides a high sensitivity and specificity for detection of fungal DNA and rapidly identifies most of clinically relevant Aspergillus species.
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Aspergilosis , Aspergillus/clasificación , Aspergillus/aislamiento & purificación , Fungemia , Reacción en Cadena de la Polimerasa/métodos , Aspergilosis/diagnóstico , Aspergilosis/microbiología , Aspergillus/genética , ADN de Hongos/análisis , ADN de Hongos/aislamiento & purificación , Fungemia/diagnóstico , Fungemia/microbiología , Humanos , Técnicas de Tipificación Micológica , Sensibilidad y Especificidad , Especificidad de la EspecieRESUMEN
Our objective was to evaluate the feasibility of a molecular assay based on a real-time PCR technique, carried out with a LightCycler instrument (Roche Biochemicals), to identify Mycobacterium tuberculosis bacilli and to detect rifampin and isoniazid resistance in DNA extracts from sputum samples. We studied three genes: rpoB, which is associated with rifampin resistance, and katG and inhA, which are associated with isoniazid resistance. A total of 205 sputum samples collected from 108 patients diagnosed with pulmonary tuberculosis with positive auramine-rhodamine-staining (AR) sputum samples, were tested. The sensitivities of the LightCycler PCR assay for the positive AR specimens was 97.5% (200 of 205) for rpoB and inhA genes and 96.5% (198 of 205) for the katG gene. For the total number of patients tested, the sensitivity was 100% (108 of 108 patients) for rifampin, whereas the sensitivity was 98.1% (106 of 108 patients) for isoniazid. Full agreement was found with the Bactec MGIT 960 method and the genotype inferred from the LightCycler data for rifampin. The phenotypic method for isoniazid reported 13 resistant strains (> or = 0.1 microg/ml). In seven (53.8%) strains there was a concordance between both methods, but we found that six (46.2%) strains reported as resistant by the phenotypic method were determined to be susceptible by real-time PCR. For the 75 strains reported as susceptible by the phenotypic method, the concordance with the LightCycler data was 100%. Our results demonstrate that rifampin-resistant M. tuberculosis could be detected in DNA extracted from auramine-rhodamine-positive sputum samples in a single-tube assay that took less than 3 h to perform for a collection of auramine-rhodamine-positive specimens obtained from patients with culture-documented pulmonary tuberculosis. Similarly, this occurs in half of the isoniazid-resistant M. tuberculosis DNA extracted from auramine-rhodamine-positive specimens.
Asunto(s)
Antituberculosos/farmacología , Farmacorresistencia Bacteriana , Isoniazida/farmacología , Mycobacterium tuberculosis/efectos de los fármacos , Reacción en Cadena de la Polimerasa/métodos , Rifampin/farmacología , Esputo/microbiología , Benzofenoneido/metabolismo , ADN Bacteriano/análisis , Genotipo , Humanos , Pruebas de Sensibilidad Microbiana , Mycobacterium tuberculosis/clasificación , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/aislamiento & purificación , Fenotipo , Rodaminas/metabolismo , Sensibilidad y Especificidad , Tuberculosis Pulmonar/microbiologíaRESUMEN
In this study we designed two pairs of probes for the detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis with real-time PCR procedures. One pair of probes spans the region between codon 510 and 528 of the rpoB gene, and the other one screens for mutation at the regulatory region of the inhA gene. We have evaluated these probes in combination with two other pairs of probes previously described to detect mutations in 20 susceptible and 53 unique resistant M. tuberculosis clinical isolates. We were able to detect nine different mutations affecting five codons of the rpoB gene, two different mutations at codon 315 of the katG gene and a nucleotide substitution (C209T) in the regulatory region of the inhA gene within two hours turnaround.