title: Recombinant dense granular protein (GRA5) for detection of human toxoplasmosis by Western blot.
abstract: Toxoplasma gondii infects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of whole Toxoplasma lysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressed T. gondii dense granular protein-5 (GRA5) in Escherichia coli and isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronic T. gondii infections (sensitivities of 46.8% and 61.2%, resp.).
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.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Yee Ling
authors.initials: YL
authors.lastname: Lau
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Mun Yik
authors.initials: MY
authors.lastname: Fong
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Veeranoot
authors.initials: V
authors.lastname: Nissapatorn
authors.affiliation: Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Hemah
authors.initials: H
authors.lastname: Andiappan
mesh_terms: D000953:Antigens, Protozoan / Q000737:chemistry* / Q000235:genetics; D015153:Blotting, Western / Q000379:methods*; D006801:Humans; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D014122:Toxoplasma; D014123:Toxoplasmosis / Q000097:blood*
chemical_list: D000953:Antigens, Protozoan; C095046:GRA5 protein, Toxoplasma gondii; D015800:Protozoan Proteins; D011994:Recombinant Proteins