1. Evaluation of liver fluke recombinant cathepsin B-1 protease as a serodiagnostic antigen for human opisthorchiasis
- Author
-
Sripa, Jittiyawadee, Brindley, Paul J., Sripa, Banchob, Loukas, Alex, Kaewkes, Sasithorn, and Laha, Thewarach
- Subjects
- *
LIVER flukes , *PEPTIDES , *RECOMBINANT proteins , *OPISTHORCHIASIS , *SERODIAGNOSIS , *PROTEOLYTIC enzymes , *CYSTEINE proteinases - Abstract
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. [Copyright &y& Elsevier]
- Published
- 2012
- Full Text
- View/download PDF