title: Evaluation of Toxoplasma gondii-recombinant dense granular protein (GRA2) for serodiagnosis by western blot.
abstract: Toxoplasma gondii infects all warm-blooded animals including humans, causing serious public health problems and great economic loss in the food industry. Commonly used serological tests involve preparation of whole Toxoplasma lysate antigens from tachyzoites which are costly and hazardous. An alternative method for better antigen production involving the prokaryotic expression system was therefore used in this study. Recombinant dense granular protein, GRA2, was successfully cloned, expressed, and purified in Escherichia coli, BL21 (DE3) pLysS. The potential of this purified antigen for diagnosis of human infections was evaluated through western blot analysis against 100 human serum samples. Results showed that the rGRA2 protein has 100 and 61.5 % sensitivity towards acute and chronic infection, respectively, in T. gondii-infected humans, indicating that this protein is useful in differentiating present and past infections. Therefore, it is suitable to be used as a sensitive and specific molecular marker for the serodiagnosis of Toxoplasma infection in both humans and animals.
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia.
authors.forename: Xiao Teng
authors.initials: XT
authors.lastname: Ching
authors.forename: Yee Ling
authors.initials: YL
authors.lastname: Lau
authors.forename: Mun Yik
authors.initials: MY
authors.lastname: Fong
authors.forename: Veeranoot
authors.initials: V
authors.lastname: Nissapatorn
mesh_terms: D000913:Antibodies, Protozoan / Q000097:blood* / Q000235:genetics; D000953:Antigens, Protozoan / Q000235:genetics; D015153:Blotting, Western / Q000379:methods*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D015800:Protozoan Proteins; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology; D014123:Toxoplasmosis / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; C106990:Gra2 protein, Toxoplasma gondii; D015800:Protozoan Proteins; D011994:Recombinant Proteins