a Fast-track Diagnostics, Esch-sur-Alzette/LU b Fondation Mérieux, Lyon/FR c Institut Pasteur, Paris/FR d ICDDR,B, Dhaka/BD e Fast-track Diagnostics, Luxembourg/LU Purpose: Enteric fever due to Salmonella remains a significant public health problem, predominantly in children in developing countries. Typhoid fever affects more than 20 million people per year associated to about 300,000 deaths. These high rates are mainly due to the lack of fast, reliable and inexpensive diagnostic assays. Diagnosis of typhoid remains a challenge because a low bacterial load can be responsible of an illness and also because the volume of blood that can be collected from children is limited.To overcome the problem of sensitivity, the first goal of this study was to define an appropriate pre-enrichment method of the collected sample before nucleic-acid extraction. The second objective was to develop a sensitive multiplex real-time PCR assay for detection of the common etiological agents of Enteric fever: Salmonella enterica spp., Salmonella typhi and Salmonella paratyphi A.Methods & Materials: To determine the best pre-enrichment medium, we spiked S. paratyphi A within blood samples, added the enrichment medium, cultured for 5 h at 37 • C, performed nucleic acid extraction and determined the yield of specific nucleic acids by real-time PCR. We tested several commercial enrichment broths and a home-made TSB medium supplemented with Oxgall (inhibitor of Gram+ bacteria). The results show that the Selenite broth (Copan) and the TSB-Oxgall medium are the most efficient pre-enrichment methods of free bacteria in blood samples.Results: In parallel we developed a multiplex real-time PCR assay that distinguish between infection by S. enterica spp., S. typhi and/or S. paratyphi A (+ internal control). This assay shows a limit of detection between 1E+02 and 1E+03 copies/ml and a linearity from 1E+09 to 1E+03 copies/ml for all pathogens. The specificity of the assay was validated with various negative samples which did not generate any positive signals. Furthermore, different positive materials containing bacteria, parasites, and viruses were evaluated with this assay and no other than the expected bacteria were detected.Conclusion:In conclusion, this study shows the development of an efficient diagnostic pipeline that combines a quick preenrichment of infected blood samples with a sensitive real-time multiplex PCR assay that could be used as a reference diagnostic method for Enteric fever infectious agents. http://dx.

Efforts to enhance standard precautions and to isolate patients with positive routine clinical cultures during 3 years were insufficient to decrease multidrug-resistant bacteria infection rates. Routine screening for carriage in high-risk patients may be necessary to halt transmission and control the hospital reservoir.

Cette étude s'intéresse aux ressources situationnelles que les élèves utilisent lors de leçons d'éducation physique en musculation conçues dans un format pédagogique en ateliers, en explorant les liens entre l'arrangement spatial des ateliers, la nature des objets matériels présents dans chaque atelier et les interactions entre les élèves. Elle a été conduite selon le cadre théorique et méthodologique du « cours d'action ». Deux types de données ont été recueillis : (a) des données d'enregistrement audio et vidéo des élèves en classe et (b) des données de verbalisations lors d'entretiens d'autoconfrontation. Les résultats révèlent chez les élèves réalisant les exercices (a) l'existence de quatre préoccupations typiques dirigées vers des ateliers distants associées à trois actions typiques dans l'activité des élèves, et (b) des relations entre ces actions typiques et le type d'appareil de musculation présent dans l'atelier. Ces résultats accréditent l'idée de la classe comme un système éco-social.

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