Characterization and purification of Parafilaria bovicola antigens by chromatofocusing to enhance specificity in serodiagnosis.
A study was conducted to determine if the purification of Parafilaria bovicola antigens can increase the specificity of serodiagnosis of parafilariasis in enzyme-linked immunosorbent assay. Antigens released from adult worms of P. bovicola were separated by chromatofocusing on a polybuffer exchanger of the pH range 7.3-4.0 Polypeptide analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis showed the presence of four major polypeptides with MWs of 41, 36, 24 and 20 kDa. Additional biochemical characterization identified the 24- and 20-kDa polypeptides as hydrophobic glycoproteins. The chromatofocusing purification procedures were also applied for separation of a whole-worm extract. Again, the 41- and 36-kDa antigens were identified in separate peak fractions. Using ELISA, it was shown that the 41- and 35-kDa antigens were recognized by bovine antibodies specific for P. bovicola, but not by other sera collected from cattle infected by Onchocerca gutturosa, Onchocerca lienalis, Ostertagia ostertagi and Dictyocaulus viviparus. The serological evaluation strongly suggests that the 41- and 36-kDa antigens are P. bovicola specific.
Sundquist
B
B
National Veterinary Institute, Uppsala, Sweden.
Bech-Nielsen
S
S
Zakrisson
G
G
D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis*; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005368:Filariasis / Q000175:diagnosis / Q000662:veterinary*; D005370:Filarioidea / Q000276:immunology*; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera / Q000276:immunology; D007525:Isoelectric Focusing
D000947:Antigens, Helminth; D007106:Immune Sera