Identification and partial purification of a 26 kilodalton antigen of adult Haemonchus contortus.
Adult H. contortus soluble extracts were fractionated by means of gel filtration (S-200) and anion exchange chromatography (DEAE-Sephacel). Fractions from both analyses were checked by ELISA and western blotting (WB) with sera from lambs infected with H. contortus, monospecific heterologous sera (anti-Trichostrongylus colubriformis and anti-Teladorsagia circumcinta) and sera from naturally infected sheep with mixed trichostrongylid infections. High cross reactivity was seen between H. contortus and heterologous sera, particularly with the anti-T. colubriformis serum, when fractions from gel filtration were checked by ELISA. Individual fractions containing the highest positive/negative and positive/heterologous ratios were pooled and analysed by SDS-PAGE. One of the pools (A4) containing 2 regions around 48-55 and 25-27 kDa were strongly recognized by homologous sera in WB. Similar results were obtained with the first peak eluted in the DEAE-Sephacel chromatography with NaCl 0.1 M. The pooled fraction A4 from gel filtration was further fractionated by anion exchange chromatography and the peak obtained with the NaCl gradient contained a ca. 26 kDa antigen apparently specific for the diagnosis of H. contortus infections in lambs.
Gomez-Muñoz
M T
MT
Departamento de Patologia Animal I, Fac. Veterinaria, Universidad Complutense de Madrid, Spain.
Cuquerella
M
M
Alunda
J M
JM
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood / Q000276:immunology; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D006188:Haemonchiasis / Q000276:immunology / Q000662:veterinary; D006189:Haemonchus / Q000276:immunology* / Q000302:isolation & purification; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000469:parasitology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth