Final Activity Report Summary - ADIPAR (Advanced diagnostics of parasitic diseases) The main focus of the 'Advanced diagnostics of parasitic diseases' (ADIPAR) project was the implementation of advanced techniques in the field of parasite diagnostics in order to increase the national and international competitiveness of the W. Stefanski institute of parasitology of the Polish academy of sciences. This goal involved the development of appropriate laboratory infrastructure and introduction of new research methods.Laboratory facilities for molecular biology techniques have been reorganised according to European standards. Work areas have been designed to enable DNA amplification to minimise the chances of contamination and therefore false results allowing consistent and repeatable DNA analysis. This was especially important due to the introduction of many new diagnostic, PCR-based assays. Furthermore, laboratory infrastructure was developed for the purpose of introducing recombinant DNA technology, an entirely new field of competence which resulted in the erection of a 'Genetically modified organism' (GMO) laboratory, approved by the ministry of the environment.Throughout the duration of the project many new diagnostic assays have been implemented. A PCR method for the detection of 'Ashwortius sidemi', an emerging Gastro-intestinal (GI) nematode was developed. This is the first molecular assay available for this parasite. Furthermore a multiplex PCR assay for the simultaneous identification of four important GI nematode species, including A. sidemi and Haemonchus contortus was elaborated. This test is able to detect all four species in a single reaction tube, which is time and cost-efficient Another multiplex PCR assay was implemented for the differentiation of four Trichinella species, which are important, life-threatening human parasites transmitted by meat. The institute is currently the only facility in Poland performing this important assay. Molecular assays for genotyping Giardia and Crytposporidium have also been introduced. These two protozoan parasites can be life threatening in immunocomprised persons. The implemented PCR-RFLP method allows the differentiation of the most important species and genotypes in a single reaction. With these assays an entirely new area of competence has been introduced into the Institute.An Iscom-ELISA method for the detection of Neospora caninum in red deer has also been developed. N. caninum is an important cause of reproductive failure in many domesticated and wild ruminants but little is known about its epidemiology. After validation, the test was used to provide unique data about the prevalence of deer infection in Poland. A series of novel research techniques have been introduced throughout the course of the project. The first was 'two dimensional electrophoresis' (2DE), a very important proteomic technique offering a detailed visual overview of parasite proteins, comparing protein profiles in search of diagnostic markers, as well as immunological studies through Western-blot analysis.Northern and Southern blotting for studies on gene expression and diagnostics of infectious diseases have also been implemented. A big group of methods introduced is connected with the new field of competence - recombinant DNA technology. They are among others: cloning of parasite genes, transformation techniques, bacterial culturing, expression of recombinant proteins, bioinformatic analysis and protein purification.An important outcome was the discovery of a novel diagnostic and/or vaccine candidate for Fasciola hepatica - cathepsin B3. There have been also many events organised during the project: two conferences, numerous seminars and workshops which contributed to the integration of the Institute into the national and international research community and increasing the awareness of society. The project also enhanced international scientific cooperation resulting in joint research projects. Overall, the project had a profound effect on the institute's development and fulfilled its scientific and non-scientific objectives.
