Multiple recombinant antigens of Clonorchis sinensis for serodiagnosis of human clonorchiasis

Abstract

Antigenic proteins from Clonorchis sinensis have been previously purified and evaluated for their antigenicity to enable the serodiagnosis of clonorchiasis. Though they were of high specificity, molecularly defined proteins were reported to be less sensitive as single antigens than crude antigen. To resolve this issue, 11 clones were selected by immunoscreening an adult C. sinensis cDNA library using infected human sera. Mixed antigens were prepared using recombinant proteins of positive clones and investigated for antigenicity by immunoblotting against C. sinensis- and helminth-infected patient sera. A mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases (Cs28GST and Cs26GST) produced 76% sensitivity and 95% specificity. Furthermore, a triple mix of recombinant Cs26GST and Cs28GST with vitelline precursor protein pushed up the sensitivity to 87% and maintained specificity at 95%. It is proposed that multiple antigen mixes should be further studied to develop rapid serodiagnostic test kits for the serodiagnosis of human clonorchiasis.

Development of an Immunochromatographic Point-of-Care Test for Serodiagnosis of Opisthorchiasis and Clonorchiasis

“…[16][17][18] In serodiagnosis of human clonorchiasis, ELISA and/or immunoblotting has also been performed using somatic antigens produced from adult C. sinensis 19,20 and recombinant C. sinensis antigen. [21][22][23][24] In general, these methods are time consuming and cannot be used at patient contact. Here, we report the development and testing of a rapid diagnostic immunochromatographic test (ICT) for serodiagnosis of human opisthorchiasis and clonorchiasis.…”

Molecular characterization of Clonorchis sinensis tetraspanin 2 extracellular loop 2

“…Cocktailed antigens can be used to overcome the limitation. Recently, multiple mixed antigen of recombinant 28 and 26 kDa glutathione Stransferases and vitelline precursor protein showed elevated diagnostic sensitivity and specificity (Li et al 2011). Bacterially produced rCsTSP2EC2 showed specific reactivity with human clonorchiasis sera by immunoblot in this study.…”

The Diagnosis and Classification of Parasitic Diseases of the Liver

“…However, an accurate and feasible diagnostic method for clonorchiasis is US that characterize clonorchiasis as diffuse, mild, uniform dilatation of the small peripheral intra-hepatic bile ducts without a focal obstructing lesion; the CT imaging of clonorchiasis is same as those observed by US [ 59 ]. Li et al [ 58 ] reported that the recombinant Cs26GST and Cs28GST proteins are specifi c serodiagnostic antigens for human clonorchiasis, because of their non-cross-reactivity to the sera of paragonimiasis, schistosomasis, or cysticercosis; thus, a mixed antigen of recombinant 28 and 26 kDa glutathion S-transferases, Cs28GST and Cs26GST (producing 76 % sensitivity and 95 % specifi city), can be considered an useful serodiagnostic reagent for human clonorchiasis. The C .…”

“…The diagnosis of clonorchiasis is done by microscopic fi ndings of fecal eggs [ 58 ], which are oval (measuring 27-35 × 12-20 μm), with an operculum at the slender end and prominent shoulders; broad abopercular end with small spine-like structure [ 56 ]. However, an accurate and feasible diagnostic method for clonorchiasis is US that characterize clonorchiasis as diffuse, mild, uniform dilatation of the small peripheral intra-hepatic bile ducts without a focal obstructing lesion; the CT imaging of clonorchiasis is same as those observed by US [ 59 ].…”