Partial purification and characterization of Ascaridia galli diagnostic worm antigen.
Partial purification of Ascaridia galli whole worm extract was conducted by Cyanogen bromide Sepharose 4B immunoaffinity column chromatography. The resulted fraction was characterized by sodium dodecyle sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing. The fraction was found to be consisted of six bands of 207 KDa, 157 KDa. 110 KDa, 103 KDa, 76 KDa and 41 KDa. This profile was compared with that of whole worm and excretory-secretory antigens. Both antigens were resolved into multiple bands in both high and low molecular weight ranges. The isoelectric focusing of the fraction displayed 8 bands of isoelectric points 7.5, 7.0, 6.8, 6.5, 6.2, 5.8. 5.3 and 4.6. The potency of this fraction in the diagnosis of natural ascaridiosis in chickens was assessed by ELISA compared with that of whole worm and ES antigens. The affinity purified fraction showed higher potentials in the diagnosis of A. galli infection in chickens than whole worm antigen at any sera dilution and than ES antigen at high sera dilutions. While ES antigen of the worms revealed higher diagnostic capabilities than whole worm extract. The current research recommends utilization of the affinity isolated fraction in the diagnosis of natural ascaridiosis in chickens.
Abdel Rahman
Eman H
EH
Department of Parasitology and Animal Diseases, National Research Center, Dokki, Giza, Egypt.
Khalil
Fathia A M
FA
D000818:Animals; D000947:Antigens, Helminth / Q000302:isolation & purification*; D001197:Ascaridia / Q000276:immunology*; D001198:Ascaridiasis / Q000175:diagnosis / Q000662:veterinary*; D002645:Chickens / Q000469:parasitology*; D002846:Chromatography, Affinity / Q000379:methods / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary; D007525:Isoelectric Focusing / Q000379:methods / Q000662:veterinary; D011201:Poultry Diseases / Q000175:diagnosis* / Q000469:parasitology
D000947:Antigens, Helminth