Improvement of Leishmania major Amastigote ELISA Test Using Various Antigen-Antibody Dissociating Agents and Pepsin Digestion
2004
Mirjalili, A., Razi Vaccine & Serum Research Institute, | Alimohammadian, M.H., Pasteur Institute of Iran, Tehran, Iran | Khabiri, A., Pasteur Institute of Iran, Tehran, Iran
Amastigotes are main pathogenic stage of Leishmania parasite in mammalian host. During the course of leishmania infection the amastigote surface would be covered by host proteins like immunoglobulins and complement components. These proteins produce false positive reaction and low signal/noise ratio in evaluation of antibody raising against lesion-derived amastigote by indirect ELISA. To overcome this problem we were treated lesion-derived amastigotes with several known antigen-antibody dissociating agents like glycine, potassium thiosufate, chloroquine, guanidine HCI, urea, etc. Our results indicated that these agents were ineffective in ELISA results and could not produce a clear discrimination between positive and negative controls. We were also studied the effect of pepsin digestion on amastigotes and coated these pepsin-treated amastigotes to check the ELISA results. Our results revealed combination of tests including pepsin treated amastigotes coated in alkaline pH and protein A-peroxidase conjugates as secondary antibody in an indirect ELISA design, possess high signal/noise ratio i.e. low background color.
Afficher plus [+] Moins [-]Mots clés AGROVOC
Informations bibliographiques
Cette notice bibliographique a été fournie par Agricultural Research and Education Organization
Découvrez la collection de ce fournisseur de données dans AGRIS