title: Evaluation of Pichia pastoris-expressed recombinant rhoptry protein 2 of Toxoplasma gondii for its application in diagnosis of toxoplasmosis.
abstract: Rhoptry protein 2 (ROP2) of Toxoplasma gondii is a rhoptry-secreted protein that plays a critical role in parasitophorous vacuole membrane formation during invasion. In previous studies, ROP2 has been shown to be efficient in triggering humoral and cell-mediated responses. High immunogenicity of ROP2 makes it a potential candidate for diagnosis and vaccination against toxoplasmosis. In this study, the ROP2 gene was cloned into pPICZα A expression vector and extracellularly expressed in the yeast Pichia pastoris, which has numerous advantages over other expression systems for eukaryotic proteins expression. The effectiveness of the secreted recombinant ROP2 as a diagnosis agent was assessed by Western Blot with 200 human serum samples. Recombinant ROP2 reacted with toxoplasmosis-positive human serum samples and yielded an overall sensitivity of 90% and specificity of 95%. However, recombinant ROP2 is a better marker for detection of IgG (91.7%) rather than IgM (80%).
authors: Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia.
authors: Phooi Yee
authors: PY
authors: Chang
authors: Mun Yik
authors: MY
authors: Fong
authors: Veeranoot
authors: V
authors: Nissapatorn
authors: Yee Ling
authors: YL
authors: Lau
mesh_terms: D015153:Blotting, Western / Q000379:methods; D003001:Cloning, Molecular; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000235:genetics / Q000378:metabolism*; D014123:Toxoplasmosis / Q000097:blood / Q000175:diagnosis*
chemical_list: D008565:Membrane Proteins; D015800:Protozoan Proteins; C090555:ROP 2 protein, Toxoplasma gondii