ABSTRACT
INTRODUCTION: Water sample culturing is the reference method to detect Legionella spp. in sanitary facilities. Until 2017, UNE-EN ISO 11731 only included the GVPC medium, which inhibits interfering microbiota but hinders the growth of Legionella spp. To improve its recovery, the new standard incorporates the BCYE medium into the working protocol. METHODS: We inoculated 1306 sanitary water samples onto BCYE and GVPC according to an accredited internal procedure. We compared the number of cfu/L of Legionella spp. detected in both media. RESULTS: The median in BCYE was 2000 cfu/L higher than in GVPC (Pâ¯=â¯.000). In the presence of high amounts of interfering microbiota, both media were similar; in the absence or low interfering microbiota BCYE was four times more sensitive than GVPC (Pâ¯=â¯.000). CONCLUSION: Including BCYE in the analysis of sanitary water significantly improves the recovery of Legionella spp. in low contaminated samples.
Subject(s)
Humans , Animals , Female , Adult , Crohn Disease/complications , Intestinal Diseases, Parasitic/complications , Myiasis/parasitology , Crohn Disease/drug therapy , Crohn Disease/parasitology , Diptera/growth & development , Disease Susceptibility , Feces/parasitology , Immunocompromised Host , Immunosuppressive Agents/therapeutic use , Intestinal Diseases, Parasitic/parasitology , Larva , Myiasis/complicationsSubject(s)
Crohn Disease/complications , Intestinal Diseases, Parasitic/complications , Myiasis/complications , Adult , Animals , Crohn Disease/drug therapy , Crohn Disease/parasitology , Diptera/growth & development , Disease Susceptibility , Feces/parasitology , Female , Humans , Immunocompromised Host , Immunosuppressive Agents/therapeutic use , Intestinal Diseases, Parasitic/parasitology , Larva , Myiasis/parasitologyABSTRACT
Se han descrito 6 genotipos y hasta 80 subtipos del virus de la hepatitis C (VHC). El conocimiento del genotipo infectante es crucial para tratar adecuadamente al paciente. El método de referencia para la tipificación molecular del VHC es la secuenciación completa del genoma viral y su posterior análisis filogenético. Sin embargo, esta posibilidad no está al alcance de la mayoría de los laboratorios hospitalarios. Existen varios métodos alternativos comercializados; los de uso más extendido se basan en el análisis de la región 5'UTR mediante amplificación y posterior secuenciación directa (TRUGENE 5'NC Genotyping Kit; Visible Genetics, Toronto, Ontario, Canadá) o hibridación inversa de los amplicones con sondas dispuestas en línea sobre membrana-LiPA(INNO-LiPA HCV I/II/ VERSANT HCV Genotype Assay 1.0; Innogenetics, N.V., Zwijnaarde, Bayer Healthcare, Bélgica). Los resultados obtenidos con estos ensayos son generalmente superponibles; identifican correctamente los genotipos en un 95-100% (clasifican algunos genotipos 4 y 6 como genotipos 1), pero su índice de acierto es menor (70-85%) con los subtipos, particularmente 1a,1b, 2a/2c y 4a/4c. Existe una nueva generación de pruebas comercializadas con mejores prestaciones: GEN-ETI-K DEIA kit (Sorin, Saluggia, Italy) que analiza la región core; TRUGENE NS5B Genotyping Assay (Bayer Healhcare, Bélgica), la región NS5B; Real Time HCV Genotyping Test (Abbott, MS) las regiones 5'UTR y la NS5B y VERSANT 2.0 (Bayer), las regiones 5'UTR y core; estos ensayos son muy certeros en la determinación del genotipo y de la mayoría de subtipos conflictivos, particularmente 1a y 1b
Six genotypes and up to 80 subtypes of hepatitis C virus (HCV) have been described. Knowledge of the infecting genotype is crucial for appropriate therapeutic management of HCV infection. Whole genome sequencing and subsequent phylogenetic analysis is the gold standard for HCV molecular typing; however, this procedure cannot be implemented in routine clinical laboratories. A number of alternative methods have been commercialized, the most widely used being those that target the 5'UTR region. In these assays, amplicons are first generated and then analyzed by direct sequencing (TRUGENE 5'NC genotyping kit; Visible Genetics, Toronto, Notario, Canada) or by reverse hybridization of amplicons with membrane immobilized line probes -LiPA- (INNO-LiPA HCV I/II/ VERSANT HCV genotype assay 1.0; Innogenetics, N.V., Zwijnaarde, Belgium/Bayer Healthcare). Both assays produce comparable results: the overall accuracy for genotype determination is 95-100% (a number of genotypes 4 and 6 are misclassified as genotype 1), but only 70-85% for subtyping (with frequent mistyping of subtypes 1a, 1b, 2a/2c, 4a/4c). A new generation of commercialized assays with better performance is currently available: GEN-ETI-K DEIA kit (Sorin, Saluggia, Italy), which analyzes the core region, TRUGENE NS5B genotyping assay (Bayer Healhcare) for the NS5B region, RealTime HCV Genotyping Test (Abbott, MS) for both 5'UTR and NS5B regions and VERSANT 2.0 (Bayer), both for the 5'UTR and core regions; these assays are highly accurate for genotype determination and typing of most difficult subtypes, particularly 1a and 1b
