Molecular cloning and identification of a novel Clonorchis sinensis gene encoding a tegumental protein.

The tegumental membrane of platyhelminth parasites is of crucial importance for modulation of the host response and parasite survival. A cDNA encoding a novel tegumental protein 20.8 kDa (TP20.8) was found by large-scale sequencing of a Clonorchis sinensis cDNA library. This new cDNA was 755 bp long containing an open reading frame of 555 bp, which encoded a 20.8-kDa protein with an isoelectric point of 4.33. The deduced amino acid sequence exhibits 40 and 37% identity to Schistosoma japonicum sj20.8 and Schistosoma mansoni Sm 20.8, respectively. TP20.8 transcripts were detected in the adult worm and metacercariae cDNA libraries of C. sinensis but not in the egg. Recombinant C. sinensis TP20.8 protein was produced and purified from Escherichia coli BL21. Using specific anti-recombinant TP20.8 protein sera, the TP20.8 protein was immunohistochemically localized to the outer-surface membrane of C. sinensis. The specificity and sensitivity of the recombinant antigen for serologic diagnosis was assessed by enzyme-linked immunosorbent assay using serum from 100 patients with clonorchiasis, 20 patients with schistosomiasis, and 30 negative controls. The sensitivity was 68%, and the specificity was 84%. The antigen was less useful for the serodiagnosis of clonorchiasis with IgG.