Journal of Mazandaran University of Medical Sciences (Jan 2008)
Designing of ELISA kit for diagnosis of anti-Toxoplasma specific IgG, using exerted/secreted antigens and its
Abstract
AbstractBackground and Purpose: Many studies report that Toxoplasma gondii excreted / secreted antigens (E/SA) appear to be a suitable marker for toxoplasmosis serodiagnosis. Most of these studies have used E/SA, obtained from supernatant of Toxoplasma cell culture, or by incubating tachyzoites in cell free media (RPMI-1640). The present study for the detection of Toxoplasma IgG in human serum was evaluated, using the components of peritoneal fluid of infected mice (as another source of E/SA). Materials and Methods: Peritoneal fluids of mice infected by interaperitoneal (IP) inoculation of Toxoplasma tachyzoites were collected after 3 days and centrifuged at 750×g for 15 minutes. Then the supernatant was precipitated with ammonium sulphate solution (40% saturated) and used as components encompassing E/SA.Forty (40) none- infected ( without anti- toxoplasma antibodies) and thirty- two (32) positive (with IgG to toxoplasma) human serum samples were selected (all sera were first tested by standard method for detection of IgG antibodies anti-T. gondii) then, the sera were tested by ELISA using E/SA.Results: The cut-off point with 95% confidences was found to be 0.78. Moreover, sensitivity and specificity of the method were determined to be 84% and 92%, respectively.Conclusion: The present results indicate that peritoneal exudates from mice infected with T.gondii, may be used as a source of anti-genic material for the detection of Toxoplasma-specific IgG. Furthermore, it may be valuable for the development of new tools in the serodiagnosis of toxoplasmosi
