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Contenido archivado el 2024-06-10

Field applicable antigen assays for human schistosomiasis: A multicentre development and implementation study

Objetivo

* field assays for detection of schistosome antigens CAA and CCA in serum and urine will be developed in reagent strip and/or filter format using colloidal gold-labelled monoclonal antibodies (Mabs).

* assays will be tested in pilot field trials in parallel with magnetic bead assays for detection of CAA and CCA in serum and urine of patients infected with the major Schistosoma species (S. mansoni, S. haematobium, S. japonicum).

* selected assays will be applied in sero-epidemiological studies, in chemotherapy follow-up, and reinfection studies (S.m. S.h.) for assessment of antigen kinetics (in relation to egg output).

* new MAbs for detection of circulating antigens will be developed for S. japonicum in particular, and tested and applied in assays as described above.
Expected Outcome

* a field applicable diagnostic assay for schistosomiasis
* a field applicable sample pre-treatment method
* a field applicable sample collection and storage method based on fibre web
* anti Schistosoma japonicum McAb cell lines applicable in specific and sensitive assays.
Using McAbs already developed and optimized for ELISA's detecting parasite antigens in serum and/or urine, the format of a lateral flow diagnostic device (cf. pregnancy test) will be evaluated and optimized. This format is based on the reaction of applied sample first passing and dissolving a dried labelled antibody, and subsequently migrating to a zone of bound capture antibody. This technology format is now becoming commercially available and the necessary contacts have already been made. This approach will result in a very rapid assay (approx. 4 minutes) but probably with a sensitivity only high enough to identify moderate and heavy infections.

Starting at the end of the first year, the field-applicable assays developed in Leiden will be evaluated by the three partners in the endemic areas in study groups of about 50 infected and 50 uninfected individuals. For these studies, the rapid ELISA will be used as a reference assay. It is anticipated that for each of the Schistosoma species this number of samples and the available reference data will allow selection of the assay formats most appropriate for addressing specific studies in larger study cohorts. As follow-up of chemotherapy will be essential for evaluating sensitivity of assays before and after chemotherapy, such studies will form an integral part of the work on each of the Schistosoma species. In all studies, a 5% sample will be randomly selected for quality control of the assays, initially to be performed in Leiden, and at a later stage in the laboratory of one of the African DC Partners (e.g. in Zimbabwe).

The assays thus developed will rapidly be transferred to DC partners for evaluation under field conditions. Technology will also be transferred to the Institute of Parasitic Diseases in Shanghai, China, to allow optimization with the anti-S.j. McAbs.

The hydrophobic, chemically inert polypropylene fibre web material, which was developed by the Dept. of Medical Microbiology and Immunology in Göteborg in collaboration with Mölnlycke AB, Sweden, as sampling material for blood containing CAA, will be further investigated. It is anticipated that the fibre web sampling format could in a later phase of the project be included for quality control studies of samples which have been tested in the field.

Sample pre-treatments, although currently already relatively simple, will be further adapted for field use and applicability to larger sample sizes.

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Coordinador

LEIDEN UNIVERSITY
Aportación de la UE
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Dirección
62,Wassenaarseweg 62
2300 RC LEIDEN
Países Bajos

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