000			02899nam a2200205 4500
005			20160523100745.0
008			160523b2015 xxu||||| |||| 00| 0 eng d
041			_aeng
082			_a2433-T
100			_aMadiha Sana (2013-VA-957) _923948
110			_cDr. Muhammad Imran Rashid _96077
245			_aIndigenous Elisa Kit For Toxoplasma Gondii: Optimization Of Antibody Detection Elisa Of Sag 1 Protein As An Antigen In Mouse Model
260			_c2015.
300			_a39p.;
502			_aToxoplasma is an apicomplexan intracellular parasite which is the cause of toxoplasmosis in man and animals. It occurs by the ingestion of oocyst from feces of cats or by eating raw meat in which cysts are present. It is the one of the major cause of encephalitis and abortion in immuno-compromised animals and humans. As it is difficult to screen out infected live animals from field, it is important to vaccine animals as well as humans for toxoplasma to prevent its transmission from animals to humans and from humans to their off springs. Cloning of surface antigen genes plays an important role in development of vaccine and for serology of T. gondii. Enzyme linked immuno-sorbant assay proves to be a significant tool to estimate the humoral response elicited against expressed recombinant protein in mice. The recombinant protein of SAG1 was collected from Molecular Parasitology Laboratory, University of Veterinary and Animal Sciences, Lahore. In the previous studies, SAG1 sequence was cloned in the expression plasmid and successfully expressed in prokaryotic expression system. In the current study, rSAG1 was quantified by using BCA protein assay through BioWORLD protein quantification kit. In another experiment, the Swiss mice were immunized with 15 μg rSAG1 protein 3 times with 2 weeks intervals. Two groups of mice were formed with five mice in each group. Sera were collected after 2 weeks of each inoculation. For performing ELISA, four different experiments were performed with different concentrations i.e. 5μg/ml, 250μg/ml and 500μg/ml with two different dilutions; 1/50 and 1/20. The O.D. values of concentrations 5μg/ml and 250 μg/ml with two dilution series of 1/20 and1/50 were not observed significant while the antigen coating concentration of 500 μg/ml with 1/50 dilution showed 1:160 titre and with 1/20 dilution showed 1: 1280 titre after the 3rd shot. The O.D values with 500 CHAPTER 6 SUMMARY SUMMARY 36 μg/ml concentration with 1/20 dilution after the 3rd shot were observed significant in the inoculated group as compared to the O.D values of un-inoculated negative group. It is suggested to carry out ELISA with purified rSAG-1 protein and to optimize ELISA to test toxoplasma infected mice.
650			_aDepartment of Parasitology _94290
700			_aDr. Haroon Akbar _94909
700			_aDr. Wasim Shehzad _98918
942			_cTH
999			_c8504 _d8503