The pork tapeworm Taenia solium is now increasingly recognized as a parasite with international health implications and the capability of spreading to non-endemic areas. The growing attention to this helminth species relates primarily to the severity of health problems associated with infection of the central nervous system with the larval stage, which causes neurocysticercosis currently classified as the most important neurologic disease of parasitic origin. In spite of the theoretical possibility for eradication of T. solium cysticercosis global eradication of this infection is still unlikely in the near future. The overall goal of the current Phase II application will be to finish the development, validate through clinical trials, and launch the manufacture of serological tests for detection of specific antibodies to circulating antigens of the larval stage of Taenia solium. The tests will utilize standardized antigenic material such as synthetic peptides, recombinant proteins and combinations thereof selected from antigens identified recently in diagnostic bands of Western blots (WB) prepared from cyst material of naturally infected pigs. Two types of tests will be developed: one will be in a standard microwell ELISA format, and the second will be a rapid non-instrumental format utilizing lateral flow principles. An innovative but already proven immune-capture assay technology will be applied in both new tests, which will emphasize the advantage of using pure synthetic and recombinant antigens. The quick and simple ELISA test will ideally meet the requirements for laboratory testing of multiple samples in sero-epidemiological surveys and community-based studies, as well as for diagnosis of cysticercosis infection in individuals. The lateral flow format with capability for whole blood testing will provide a convenient and more cost-effective alternative to the current WB test, and will be suitable for small batch or individual testing in labs or field conditions. Methodological innovations that will be implemented in the lateral flow test will make its sensitivity equal to or even higher than the sensitivity of the microplate ELISA version. Both ELISA and lateral flow tests will be able to detect antibodies in samples from two hosts of T. solium, humans and pigs. Phase II of this project includes finalizing the analysis of synthetic and recombinant antigens in order to select combination(s) with a minimal number of antigens providing equal or better sensitivity than the current WB test for detection of antibodies in patients with various stages of disease, with comparably high specificity. The second major goal will be validation of technology for preparation of antigenic material and diagnostic reagents including launching the manufacture of pilot lots of the new tests. Significant efforts at this stage will be directed towards optimization and manufacture of highly sensitive lateral flow versions of the serological tests. The objective of the present project is the development of the two types of a commercial serological tests for diagnosis of infection caused by larva stage of pork tapeworm Taenia solium in human and pigs utilizing standardized antigenic material such as synthetic peptides and recombinant proteins. The first test will be in simple microwell ELISA format well suited for testing of any number of samples, using semi-automated or automated equipment in clinical labs and equipped facilities. The second test will be in rapid non-instrumental format utilizing lateral flow principles for application in field conditions or in laboratories for testing relatively small batches of samples. The new tests will replace the expensive WB technique that requires specially trained personnel, and will make possible a level of standardization and accuracy in clinical and reference laboratories that is currently unattainable. The primary application of these tests will be the diagnosis of neurocysticercosis in humans, pig cysticercosis, seroepidemiological surveys and community-based studies. ? ? ?
