title: Evaluation of liver fluke recombinant cathepsin B-1 protease as a serodiagnostic antigen for human opisthorchiasis.
abstract: A cathepsin B-like cysteine protease belonging to family C1 is abundantly expressed in the transcriptome and proteome of the carcinogenic liver fluke of humans, Opisthorchis viverrini. This enzyme is present in excretory/secretory (ES) products released by parasites cultured in vitro. This study evaluated the performance of recombinant O. viverrini cathepsin B1 (rOv-CB-1) as an antigen for immunodiagnosis of opisthorchiasis. The full length Ov-CB-1 cDNA was cloned and recombinant protein was produced in catalytically active form in Pichia pastoris. The recombinant Ov-CB-1 (rOv-CB-1) was affinity purified via nickel-NTA chromatography and tested in enzyme-linked immunosorbent assays (ELISA) with human sera from an opisthorchiasis endemic area. Sera from egg-positive O. viverrini infections produced a strong IgG antibody response to rOv-CB-1 both in ELISA and immunoblot analysis. The sensitivity and specificity of the ELISA test was 67% and 81%, respectively. These findings support the feasibility of using recombinant Ov-CB-1 in ELISA for the serodiagnosis of human opisthorchiasis.
authors.affiliation: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Jittiyawadee
authors.initials: J
authors.lastname: Sripa
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Brindley
authors.forename: Banchob
authors.initials: B
authors.lastname: Sripa
authors.forename: Alex
authors.initials: A
authors.lastname: Loukas
authors.forename: Sasithorn
authors.initials: S
authors.lastname: Kaewkes
authors.forename: Thewarach
authors.initials: T
authors.lastname: Laha
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood*; D000947:Antigens, Helminth / Q000097:blood / Q000235:genetics / Q000378:metabolism; D002401:Cathepsin B / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D015801:Helminth Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D008297:Male; D009889:Opisthorchiasis / Q000175:diagnosis* / Q000276:immunology / Q000469:parasitology; D009891:Opisthorchis / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D010843:Pichia; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D013785:Thailand
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D011994:Recombinant Proteins; D002401:Cathepsin B