Purification of the Clonorchis Antigen using a Diethylaminoethyl-Sephadex A50 Column

ABSTRACT

Antigens for the diagnosis of clonorchiasis have been isolated from the adult Clonorchis. Following extracttion with a phosphate-citrate buffer solution, ultrasonic disintegration, and defatting with a chloroform-methanol mixture, the antigens were fractionated in a Diethylaminoethyl-Sephadex (DEAE-Sephadex) A 50 column using sodium chloride eluates. The antigens were evaluated by carrying out both skin tests and stool examinations to determine their sensitivity, specificity and cross reaction. These DEAE fractionated antigens show less cross reaction than Melcher's antigen. DEAE fractions were protein(280mmu peak) and nucleoprotein (260mmu peak) as indicated by spectrophotometry. These purified fractions represented by the 260mmu and 280mmu peaks were not free of cross reactions nor were they completely specific for clonorchiasis. It is therefore suggested that further attempts be made to eliminate cross reactions and to enhance specificity by a more detailed and complete method of purification or of preparation of the chemical components.