Cystic echinococcosus (CE) is a serious public health problem in sheep-raising regions. Detection of E. granulosus antigens is a better immunodiagnostic tool than determination of the antibody level. The circulating protoscolex antigen (CPA) used was obtained from lung and liver cysts of sheep and camel and injected in rabbits to raise specific polyclonal antibodies (pAb) against E. granulosus. A novel immunomagnetic bead ELISA based on immunoglobulin G (IgG) for detection of CPA in sera of rabbit infected with E. granulosus was developed. Detection of CPA in serum of human patients by sandwich ELISA gave a sensitivity of 90.48% and a specificity of 91.3% compared to other parasitic infections group and 100% compared to healthy control group. On the other hand, detection of CPA in serum of human patients by sandwich ELISA with paramagnetic nanoparticles gave a sensitivity of 95.2% and a specificity of 95.5% compared to other parasitic infections group and 100% compared to healthy control group. In conclusion, sandwich ELISA with paramagnetic nanoparticles techniques appears to be sufficiently sensitive assays for the detection of human echinococcosis antigen and valuable to judge the efficacy of chemotherapy than sandwich ELISA.

Using of paramagnetic nanoparticales with immunomagnetic bead ELISA in diagnosis of hydatidosis (Echinococcus Granulosus)

Objectives : Snail-schistosome interactions in relation to immunological, and biochemical changes induced in the host’s tissues by the developing intramolluscan stages of the parasite form an integral part in  understanding the biology of infection. This study focused on determining whether there are induced proteins in an infected snail haemolymph and to determine the cross reactivity of the proteins with antibodies raised against the cercariae and worm antigens. Methodology and results : Proteomic analysis was carried out to analyze differentially expressed proteins. This was done by separation of proteins by SDS-PAGE and 2D electrophoresis on infected snail haemolymph. Later western blotting was done to check for cross reactivity of induced proteins and antibodies. The protein profiles in SDS-PAGE revealed a complex mixture of polypeptides some of which were induced or suppressed on specific days during the infection period. Separation of haemolymph  proteins by 2D electrophoresis analysis revealed a progressive increase of expressed proteins during the parasite’s developmental period in the snail, however there was no specific trend in distribution of the acidic or basic proteins. Finally, haemolymph proteins from infected snails and the control (uninfected) snails were not recognized by antibodies raised against S. mansoni antigens namely Soluble Worm Antigen (SWAP) and Soluble Cercariae Antigen (SCA). Conclusions and application of findings : Snail- schistosome interaction leads to suppression and induction of proteins. This is important in  understanding stage specific interactions of the parasite during  intramolluscan development. Lack of cross reactivity was an indication that these proteins could not be used as immunogens for vaccine design. Key words : Schistosomiasis, Biomphalaria pfeifferi, haemolymph, induced proteins

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Detection of proteins induced in the haemolymph of Biomphalaria pfeifferi infected with Schistosoma mansoni

The reaction of the haemolymph and the tissue of infected intermediate hosts, Biomphalaria alexandrina and Bulinus truncatus to Schistosoma mansoni and S. haematobium antigens were investigated using the indirect immunoperoxidase technique. A new technique, Agarose cell block was used in collection of haemolymph which helped in collecting plenty of well formed cells in comparison to the ordinary one using the cytospin. Collected haemolymph and prepared tissues of uninfected and infected B. alexandria and B. truncatus were fixed and then reacted with anti-S. mansoni and anti-S. haematobium IgG polyclonal antibodies. The haemolymph and tissue of infected B. alexandrina and B. truncatus gave a positive peroxidase reaction represented by a brown colour. In haemolymph, the positive peroxidase reaction was detected mainly in the cytoplasm of the amoebocytes. In the tissue, it was detected in epithelial cells lining the tubules, male cells in the lumen of the tubules and in female oogonia cells along the periphery of the tubules. The similarity in the strength and distribution of positive reaction in B. alexandrina and B. truncates was observed as compared to control. Thus, the immunoperoxidase technique proved to be an effective indicator for the schistosome-antigen in the snails.

Ibrahim Rabia

Papers

Immunolocalization of Schistosoma mansoni and Schistosoma haematobium antigens reacting with their Egyptian snail vectors.