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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.
AuthorsJittiyawadee Sripa, Paul J Brindley, Banchob Sripa, Alex Loukas, Sasithorn Kaewkes, Thewarach Laha
JournalParasitology international (Parasitol Int) Vol. 61 Issue 1 Pg. 191-5 (Mar 2012) ISSN: 1873-0329 [Electronic] Netherlands
PMID21704728 (Publication Type: Evaluation Study, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2011 Elsevier Ireland Ltd. All rights reserved.
Chemical References
  • Antibodies, Helminth
  • Antigens, Helminth
  • Helminth Proteins
  • Recombinant Proteins
  • Cathepsin B
Topics
  • Animals
  • Antibodies, Helminth (blood)
  • Antigens, Helminth (blood, genetics, metabolism)
  • Cathepsin B (genetics, immunology, metabolism)
  • Chromatography, Affinity
  • Enzyme-Linked Immunosorbent Assay (methods)
  • Female
  • Helminth Proteins (genetics, immunology, metabolism)
  • Humans
  • Immunoblotting
  • Male
  • Opisthorchiasis (diagnosis, immunology, parasitology)
  • Opisthorchis (enzymology, genetics, growth & development, immunology)
  • Pichia
  • Recombinant Proteins (genetics, immunology, metabolism)
  • Sensitivity and Specificity
  • Serologic Tests (methods)
  • Thailand

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