Core B will centralize the management of all of the immunology and molecular biology procedures neededto support the two scientific projects that form the Peru TMRC, thus optimizing the use of resources and maintaining rigorousquality control standards.
The specific aims of Core B are to (1) coordinate all work related to immunology and molecular biology procedures between both Research Projects and the Epidemiology/GIS/Data Management core of the Peru TMRC; (2) coordinate and oversee collection and handling procedures for all biological specimens; (3) standardize, perform, and monitor all immunological and molecular biology procedures; (4) optimize the use of laboratory personnel, equipment and supplies; (5) coordinate the register and collation of assay results; (5) provide the means for an efficient transfer and diffusion of acquired technology; (6) provide the environment for immunology and parasitology training for University students and visiting investigators. The major procedures consist of serologic and parasitologic assays for the diagnosis of T.cruzi infection in humans, guinea pigs and dogs (epimastigote antigen ELISA and immunofluoresecent antibody tests, xenodiagnosis, hemoculture), the use of Trypomastigote excreted-secreted antigens (TESA) in standard Western blot, ELISA, Falcon assay screening test - ELISA and a novel field-friendly rapid TESA-blot using pre-prepared strips to allow rapid turn- around, a range of T.cruzi molecular techniques (Detection PCR targeting nuclear DNA satellite repeats or kinetoplast DNA minicircles, Nuclear spliced leader array for classifying T. cruzi discrete typing units' and distinguishing it from T.ranglei, T.cruzi DNA RAPDS), microsatellite loci for Triatoma infestans and Surface-enhanced laser desorption ionization-time of flight mass spectrometry for the indentification of biomarkers. All point-of-care immunological tests, FAST- ELISA and the rapid field-friendly format of the TESA blot will be conducted in our field sites, Arequipa and Santa Cruz, Bolivia. PCRand RT- PCR will also be performed at the UPCH laboratory and then the technique will be transferred to Bolivia. New techniques such as strain typing using multi - locus typing will be developed at UCLA andthen transferred to the UPCH laboratory. Microsatellite loci testing for T. infestans will be performed at the CDC Division of Parasitic Diseases entomology laboratory. Hemoculture of blood from humans, animals and T. infestans feces, and xenodiagnosis in guinea pigs, will all be performed on site either in Arequipa or Santa Cruz. All proteomics work will be conducted at the Imperial College, London, UK. The implementation of these procedures will support the scientific work of the two Research Projects and provide significant training opportunities for Peruvian scientists through interchanges with the US, UK and Brazilian collaborators and consultants. PERFORMANCE.
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