title: Recombinant proteins from new constructs of SAG1 and GRA7 sequences and their usefulness to detect acute toxoplasmosis.
abstract: In this study we have cloned unreported gene fragments of Toxoplasma gondii   GRA7 and SAG1 and expressed the corresponding recombinant proteins, followed by evaluation   of their usefulness for the serological diagnosis of toxoplasmosis. Both recombinant proteins   were expressed efficiently in insoluble form, purified by single step Ni-NTA affinity   chromatography and their antigenicity to detect toxoplasma specific IgG antibodies were   determined by immunoblotting. A total of 60 serum samples from three groups of individuals   based on their anti-toxoplasma antibody profiles were tested, namely (I) IgM+, IgG+ (n=20),   (II) IgM-, IgG+ (n=20) and (III) IgM-, IgG- (n=20). Both recombinant proteins exhibited high   sensitivity (100%) with sera from Group I. rGRA7 and rSAG1 reacted 40% and 80% respectively   with Group II sera. The specificity of the recombinant proteins based on reactivities with   Group III sera were 100% and 80% with rGRA7 and rSAG1 respectively. Thus rGRA7 was found   to be better at discriminating probable acute from chronic phases of toxoplasmosis, and it   also showed higher specificity.
authors.affiliation: Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, 11800 Penang, Malaysia.
authors.forename: D
authors.initials: D
authors.lastname: Kotresha
authors.forename: D
authors.initials: D
authors.lastname: Poonam
authors.forename: Y
authors.initials: Y
authors.lastname: Muhammad Hafiznur
authors.forename: G
authors.initials: G
authors.lastname: Saadatnia
authors.forename: O
authors.initials: O
authors.lastname: Nurulhasanah
authors.forename: O
authors.initials: O
authors.lastname: Sabariah
authors.forename: S Y
authors.initials: SY
authors.lastname: Tan
authors.forename: A K
authors.initials: AK
authors.lastname: Izzati Zahidah
authors.forename: N
authors.initials: N
authors.lastname: Rahmah
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics / Q000302:isolation & purification; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D015800:Protozoan Proteins / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods; D014122:Toxoplasma / Q000235:genetics; D014123:Toxoplasmosis / Q000175:diagnosis*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; C112573:GRA7 protein, Toxoplasma gondii; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C047168:SAG1 antigen, Toxoplasma