Neisseria gonorrhoeae antimicrobial resistance in Spain: a prospective multicentre study.

OBJECTIVES: Gonococcal infection is one of the most reported sexually transmitted infections and antimicrobial resistance in Neisseria gonorrhoeae (NG) is challenging for the treatment of this infection. This observational study aimed to describe antimicrobial resistance of NG and epidemiological data from patients with gonococcal infection in eight regions of Spain, for updating the local therapeutic guidelines. METHODS: MICs of penicillin, cefixime, ceftriaxone, azithromycin, ciprofloxacin, fosfomycin and gentamicin were determined by Etest for all NG isolates recovered from 1 April 2018 to 30 September 2019 from 10 hospitals in Spain. Resistance determinants were identified using logistic regression analysis. Differences with a P value <0.05 were considered statistically significant. RESULTS: Antimicrobial susceptibility testing was performed for 2571 gonococci isolated from 2429 patients. 44.5% (945/2124) of patients were MSM. The resistance rate to extended-spectrum cephalosporins was low, with 0.2% (6/2561) of isolates resistant to ceftriaxone and 1.7% (44/2517) of isolates resistant to cefixime. The overall azithromycin resistance rate was 12.1% (310/2560), but differed greatly depending on the area. 56.2% (1366/2429) of the strains studied were ciprofloxacin resistant. MIC50 and MIC90 values of gentamicin and fosfomycin were 4 and 8 mg/L and 24 and 48 mg/L, respectively. CONCLUSIONS: Our study shows that NG susceptibility to extended-spectrum cephalosporins remains high in Spain. The azithromycin resistance rate questions the suitability of dual therapy. This study provides data of interest for updating the national treatment guidelines and highlights the need to develop and implement a national sentinel gonococcal antimicrobial susceptibility programme.

Capillary electrophoresis of PCR fragments with 5´-labelled primers for testing the SARS-Cov-2.

Due to the huge demand for SARS-Cov-2 determination,alternatives to the standard qtPCRtestsare potentially useful for increasing the number of samples screened. Our aim was to develop a direct fluorescent PCR capillary-electrophoresis detection of the viral genome. We validated this approach on several SARS-Cov-2 positive and negative samples.We isolated the naso-pharingealRNA from 20 positive and 10 negative samples. The cDNA was synthesised and two fragments of the SARS-Cov-2 were amplified. One of the primers for each pair was 5´-end fluorochrome labelled. The amplifications were subjected to capillary electrophoresis in ABI3130 sequencers to visualize the fluorescent peaks.The two SARS-Cov-2 fragments were successfully amplified in the positive samples, while the negative samples did not render fluorescent peaks. In conclusion, we describe and alternative method to identify the SARS-Cov-2 genome that could be scaled to the analysis of approximately 100 samples in less than 5 h. By combining a standard PCR with capillary electrophoresis our approach would overcome the limits imposed to many labs by the qtPCR and increase the testing capacity.

Diagnóstico de meningitis/encefalitis en UCI con sistema de PCR múltiple. ¿Es tiempo de cambio? / Meningitis/Encephalitis diagnosis in ICU using Multiplex PCR system: is it time of change?

OBJETIVO: Evaluar el impacto clínico de la PCR-múltiple FilmArray(R) panel Meningitis/Encefalitis en el diagnóstico de infecciones del sistema nervioso central y comparar los resultados obtenidos y el tiempo necesario hasta el diagnóstico con las técnicas microbiológicas convencionales. PACIENTES Y MÉTODOS: Estudio prospectivo observacional en una Unidad de Cuidados Intensivos (UCI) de adultos de un hospital de tercer nivel. Se realizó punción lumbar a todos los pacientes y en el LCR extraído se realizó FilmArray(R) panel de meningitis /encefalitis, estudio citoquímico, Gram y cultivos microbiológicos convencionales. RESULTADOS: 21 pacientes ingresados con sospecha de Meningitis/Encefalitis. Edad: mediana 58,4 años (RIQ 38,1-67,3), APACHE II: mediana 18 (RIQ 12-24). La mediana de estancia en UCI fue de 4 días (RIQ 2-6) y la hospitalaria de 17 días (RIQ 14-28). Mortalidad 14,3%. Se estableció un diagnóstico clínico final de Meningitis/Encefalitis en 16 pacientes, con diagnóstico etiológico en 12 casos (75%). La etiología más frecuente fue Streptococcus pneumoniae (8 casos). FilmArray(R) permitió diagnóstico etiológico en 3 casos con cultivo negativo y el resultado implicó cambios en el tratamiento antibiótico de 7 de los 16 pacientes (43,8%). Para la totalidad de pacientes, FilmArray(R) presentó una sensibilidad y especificidad del 100% y 90% respectivamente. La mediana de tiempo hasta la obtención del resultado de FilmArray(R) fue de 2,9 horas (RIQ 2,1-3,8) y del cultivo incluyendo antibiograma 45,1 horas (RIQ 38,9-58,7). CONCLUSIONES: FilmArray(R) panel Meningitis/Encefalitis realiza un diagnóstico etiológico más precoz que los cultivos convencionales, muestra una mayor sensibilidad y permite realizar un tratamiento antimicrobiano dirigido

OBJECTIVE: To evaluate the clinical impact of Meningitis/Encephalitis FilmArray(R) panel for the diagnosis of cerebral nervous system infection and to compare the results (including time for diagnosis) with those obtained by conventional microbiological techniques. PATIENTS AND METHODS: A prospective observational study in an Intensive Care Unit of adults from a tertiary hospital was carried out. Cerebrospinal fluid from all patients was taken by lumbar puncture and assessed by the meningitis/encephalitis FilmArray(R) panel ME, cytochemical study, Gram, and conventional microbiological cultures. RESULTS: A total of 21 patients admitted with suspicion of Meningitis/Encephalitis. Median age of patients was 58.4 years (RIQ 38.1-67.3), median APACHE II 18 (RIQ 12-24). Median stay in ICU and median hospital stay was 4 (RIQ 2-6) and 17 days (RIQ 14-28), respectively. The overall mortality was 14.3%. A final clinical diagnosis of meningitis or encephalitis was established in 16 patients, obtaining the etiological diagnosis in 12 of them (75%). The most frequent etiology was Streptococcus pneumoniae (8 cases). FilmArray(R) allowed etiological diagnosis in 3 cases in which the culture had been negative, and the results led to changes in the empirical antimicrobial therapy in 7 of 16 cases (43.8%). FilmArray(R) yielded a global sensitivity and specificity of 100% and 90%, respectively. The median time to obtain results from the latter and conventional culture (including antibiogram) was 2.9 hours (RIQ 2.1-3.8) and 45.1 hours (RIQ 38.9-58.7), respectively. CONCLUSIONS: The Meningitis/Encephalitis FilmArray(R) panel was able to establish the etiologic diagnosis faster than conventional methods. Also, it achieved a better sensitivity and led to prompt targeted antimicrobial therapy

New clinical and seasonal evidence of infections by Human Parainfluenzavirus.

Human Parainfluenzaviruses (PIVs) account for a significant proportion of viral acute respiratory infections (ARIs) in children, and are also associated with morbidity and mortality in adults, including nosocomial infections. This work aims to describe PIV genotypes and their clinical and epidemiological distribution. Between December 2016 and December 2017, 6121 samples were collected, and submitted to viral culture and genomic quantification, specifically Parainfluenza 1-4 (PIV1-4), Influenza A and B, Respiratory Syncytial Virus (RSV) A and B, Adenovirus, Metapneumovirus, Coronavirus, Rhinovirus, and Enterovirus. Normalized viral load, as (log10) copies/103 cells, was calculated as virus Ct, determined by multiple qRT-PCR, as a function of the Ct of ß-globin. PIV was confirmed in 268 cases (4.37%), and linked to both upper and lower respiratory tract disease, being more frequent in children than in adults (5.23 and 2.43%, respectively). PIV1 and PIV3 were most common (31 and 32.5%, of total PIV positive samples, respectively), with distribution being similar in children and adults, as was viral load. PIV type was correlated with seasonality: PIV3 being more frequent in winter and spring, PIV1 in summer, and PIV 4 in fall. No correlation between vial load and clinical severity was found. Novel findings were that PIV viral load was higher in fall than in other seasons, and PIV4, classically linked to mild respiratory symptoms, was circulating, in children and adults, at all levels of symptoms throughout the year.

Infección urinaria por Lelliottia amnigena (Enterobacter amnigenus): un patógeno infrecuente / No disponible

No disponible