Validation and Search of the Ideal Cut-Off of the Sysmex UF-1000i (R) Flow Cytometer for the Diagnosis of Urinary Tract Infection in a Tertiary Hospital in Spain

Urinary tract infections (UTI) are one of the most prevalent infections. A rapid and reliable screening method is useful to screen out negative samples. The objective of this study was to validate the Sysmex flow cytometer UF-1000i by evaluating its accuracy, linearity and carry-over; and define an optimal cut-off value to be used in routine practice in our hospital. For the validation of the UF-1000i cytometer, precision, linearity and carry-over were studied in samples with different counts of bacteria, leukocytes and erythrocytes. Between March and June 2016, urine samples were tested in the Clinical Microbiology Laboratory at University Miguel Servet Hospital, in Spain. Samples were analyzed with the Sysmex UF-1000i cytometer, and cultured. Growth of >= 10(5) CFUs/mL was considered positive. The validation study reveals that the precision in all the variables is acceptable; that there is a good linearity in the dilutions performed, obtaining values almost identical to those theoretically expected; and for the carry-over has practically null values. A total of 1, 220 urine specimens were included, of which 213 (17.4%) were culture positive. The optimal cut-off point of the bacteria-leukocyte combination was 138.8 bacteria or 119.8 leukocytes with an S and E of 95.3 and 70.4%, respectively. The UF-1000i cytometer is a valuable method to screen urine samples to effectively rule out UTI and, may contribute to the reduction of unnecessary urine cultures

In Vivo IS6110 profile changes in a Mycobacterium tuberculosis strain as determined by tracking over 14 years

Transposition and homologous recombination of IS6110 appear in Mycobacterium tuberculosis along in vivo sequential infec- tions. These events were checked in different clones of a successful strain, M. tuberculosis Zaragoza, with the focus on a variant in which integration of a copy of IS6110 in the origin of replication (oriC) region occurred

Comparing Two Automated Techniques for the Primary Screening-Out of Urine Culture

Urinary tract infection is the most common human infection with a high morbidity. In primary care and hospital services, conventional urine culture is a key part of infection diagnosis but results take at least 24 h. Therefore, a rapid and reliable screening method is still needed to discard negative samples as quickly as possible and to reduce the laboratory workload. In this aspect, this study aims to compare the diagnostic performance between Sysmex OF-1000i and FUS200 systems in comparison to urine culture as the gold standard. From March to June 2016, 1, 220 urine samples collected at the clinical microbiology laboratory of the "Miguel Servet" hospital were studied in parallel with both analysers, and some technical features were evaluated to select the ideal equipment. The most balanced cut-off values taking into account bacteria or leukocyte counts were 138 bacteria/mu L or 119.8 leukocyte/pl for the OF-1000i (95.3% SE and 70.4% SP), and 5.7 bacteria/mu L or 4.3 leukocyte/mu L for the FUS200 (95.8% SE and 44.4% SP). The reduction of cultured plates was 37.4% with the FUS200 and 58.3% with the UF-1000i. This study shows that both techniques improve the workflow in the laboratory, but the OF-1000i has the highest specificity at any sensitivity and the FUS200 needs a shorter processing time

Idoneidad del uso del MALDI-TOF MS para la identificación de Staphylococcus aureus y miembros del grupo de Staphylococcus intermedius (S.I.G.)

A pesar de que el género Staphylococcus es comúnmente aislado en humanos y animales, su identificación sigue siendo problemática. Este estudio evalúa la idoneidad del MALDI-TOF MS (Biotyper 3) para su identificación, comparando los resultados obtenidos mediante diferentes métodos: fenotípicos, MALDITOF MS (añadiendo o no ácido fórmico) y moleculares. Una colección de 37 cepas fue identificada por técnicas convencionales como S. aureus (n= 7), S. intermedius (n=1) y S. pseudintermedius (n=29). Los aislamientos provenían de perros, tanto sanos como enfermos (n=27), y humanos (n=10), a partir de diferentes muestras biológicas. Todas ellas fueron también identificadas por biología molecular y los cultivos puros fueron procesados en el MALDI-TOF MS. La información fue analizada con DAG_Stat. La sensibilidad, especificidad, eficiencia y el índice kappa se estimaron para cada especie bacteriana con un nivel de confianza del 95%, tomando la biología molecular como gold standard. Se detectaron Estafilococos de 27 perros y de sus dueños. Solamente una cepa de S. intermedius fue aislada, así que los parámetros relacionados con ella tienen que ser considerados con cautela. Todos los S. aureus fueron identificados correctamente. Usando MALDI-TOF MS con ácido fórmico, hubo una concordancia casi perfecta entre pruebas en la identificación de S. aureus y S. pseudintermedius. Cuando no se añadía ácido fórmico, la concordancia fue perfecta para S. aureus, mientras que para S. pseudintermedius fue buena. Este trabajo demuestra la validez, la utilidad y la fiabilidad del MALDI-TOF MS para la identificación de aislamientos bacterianos pertenecientes a Staphylococcus spp

Successful treatment of chromoblastomycosis of 36 years duration caused by Fonsecaea monophora

We report a case of chromoblastomycosis in a 67-year-old female farmer, which involved a large (20 × 30 cm) cicatricial erythematous plaque on the inner side of her right thigh. The lesion was initially a small nodule which gradually extended over 36 years. Direct microscopic examination revealed a granulomatous lesion with muriform cells surrounded by giant cells. The mould recovered in cultures was dark olivaceous and identified as Fonsecaea monophora by ribosomal internal transcribe spacer (ITS) sequence data. The lesion was successfully cured after 4 months treatment with itraconazole, but there was a relapse

Mapa de resistencias bacterianas en Atención Primaria (2010). Una herramienta esencial para el uso racional de antibióticos

Objectives:Gather and reproduce the information issued on the map of bacterial 2010 resistance for patients under the age of 15 primary care conducted by the service of Microbiology of the Hospital Universitario Miguel Servet in Zaragoza. Methods:Document establishes bacteria most frequently isolated in different organic samples (mainly stool, urine, and faringoamigdalar, otic and conjunctival exudate) as well as the percentages of resistance to antimicrobials.The results are set out in number/absolute or percentage. Results:They are offered in different tables for each germ.The most relevant data are prominent in the chapter on comments. Conclusions:Knowledge of the local resistance data facilitates the rational use of medicines.The possibility to get to know these data on an annual basis will make it possible to monitor the evolution of resistance to pathogens most prevalent in Pediatrics.Objetivos:Recopilar y reproducir la información emitida en el Mapa de Resistencias Bacterianas 2010 para pacientes menores de 15 años de Atención Primaria realizado por el Servicio de Microbiología del Hospital Universitario Miguel Servet de Zaragoza. Métodos:El documento establece los gérmenes más frecuentemente aislados en las diferentes muestras orgánicas (principalmente heces, orina, y exudados faringoamigdalar, ótico y conjuntival), así como los porcentajes de resistencias a antimicrobianos. Los resultados se exponen en números absolutos y/o porcentajes. Resultados:Son ofrecidos en tablas diferentes para cada germen. Los datos más relevantes son destacados en el capítulo de comentarios. Conclusiones:El conocimiento de los datos de resistencia locales facilita el uso racional de medicamentos. La posibilidad de ir conociendo estos datos con periodicidad anual permitirá monitorizar la evolución de las resistencias a los patógenos más prevalentes en pediatría

Acquisition of carbapenem resistance in multiresistant Klebsiella pneumoniae strains harbouring bla CTX-M-15, qnrS1 and aac(6')-Ib-cr genes

Three closely related Klebsiella pneumoniae strains isolated from the same patient harboured bla CTX-M-15, bla OXA-1, bla SHV-11, qnrS1, aac(69)-Ib-cr, oqxAB, aac(3)-II and aph(39)-Ia genes. Two of the isolates were recovered after treatment with meropenem and showed resistance to carbapenems. Sequencing of ompK35 and ompK36 porin genes of the carbapenem-resistant strains revealed the presence of premature stop codons in both, and OmpK35 and OmpK36 porins were not detected by SDS-PAGE. One carbepenem-resistant strain showed a high amount of LamB protein and did not express OmpK26 porin whereas the other strain expressed OmpK26 but not LamB. The lack of major porins apparently causes changes in the expression of other, specific, porins. © 2012 SGM