title: A rapid diagnostic test for toxoplasmosis using recombinant antigenic N-terminal half of SAG1 linked with intrinsically unstructured domain of gra2 protein.
abstract: Toxoplasma gondii is an apicomplexan parasite with a broad host range of most warm-blooded mammals including humans, of which one-thirds of the human population has been infected worldwide which can cause congenital defects, abortion, and neonatal complications. Here, we developed a rapid diagnostic test (RDT) for T. gondii infection. Antigenic N-terminal half of the major surface antigen (SAG1) was linked with intrinsically unstructured domain (IUD) of dense granule protein 2 (GRA2). The recombinant GST-GRA2-SAG1A protein was successfully expressed and purified as 51 kDa of molecular weight. Furthermore, antigenicity and solubility of the rGST-GRA2-SAG1A protein were significantly increased. The overall specificity and sensitivity of GST-GRA2-SAG1A loaded RDT (TgRDT) were estimated as 100% and 97.1% by comparing with ELISA result which uses T. gondii whole cell lysates as the antigen. The TgRDT tested with Uganda people sera for field trial and showed 31.9% of seroprevalence against T. gondii antibody. The TgRDT is proved to be a kit for rapid and easy to use with high accuracy, which would be a suitable serodiagnostic tool for toxoplasmosis.
authors: Department of Parasitology, Catholic Institute of Parasitic Disease, College of Medicine, Catholic University of Korea, Seoul 137-701, Korea.
authors: Kyoung Ju
authors: KJ
authors: Song
authors: Zhaoshou
authors: Z
authors: Yang
authors: Chom-Kyu
authors: CK
authors: Chong
authors: Jin-Soo
authors: JS
authors: Kim
authors: Kyung Chan
authors: KC
authors: Lee
authors: Tong-Soo
authors: TS
authors: Kim
authors: Ho-Woo
authors: HW
authors: Nam
mesh_terms: D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000235:genetics / Q000276:immunology*; D002648:Child; D002675:Child, Preschool; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins; D015203:Reproducibility of Results; D056910:Republic of Korea / Q000453:epidemiology; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D013997:Time Factors; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D014123:Toxoplasmosis / Q000175:diagnosis* / Q000453:epidemiology / Q000469:parasitology; D014454:Uganda / Q000453:epidemiology; D055815:Young Adult
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; C106990:Gra2 protein, Toxoplasma gondii; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C047168:SAG1 antigen, Toxoplasma
keywords: ELISA; Toxoplasma gondii; intrinsically unstructured domain; rapid diagnostic test; recombinant GST-GRA2-SAG1A; serodiagnosis