ELISA antibody determination in patients with anisakiosis or toxocariosis using affinity chromatography purified antigen.
One of the fundamental aspects of a parasitic infection diagnosis is the use of adequate antigens to develop specific and sensitive immunoassays. This fact is especially complicated in nematode infection cases because of the high cross-reactivity among different parasites in this group. We performed an evaluation of Anisakis simplex antigens purified by affinity chromatography. We used sera from 38 patients diagnosed with Anisakis sensitization and sera from 35 patients with clinical suspicion of visceral larva migrans (VLM). These sera were assayed by the ELISA method against the crude extracts (CEs) and the purified antigens. When the sera from patients diagnosed with Anisakis sensitization were tested against the A. simplex CE, the IgG was the most abundant immunoglobulin. When the A. simplex larval antigens were purified using a column of IgG anti-A. simplex (PAK) or a column of IgG anti-Ascaris suum (PAS) were tested, we observed a higher diminution in the IgG levels, which coincides with the augmentation of the mean values against the "eluted of Ascaris" (EAS antigen). When the IgE was detected, only 18.4% of the sera reacted with the PAS antigen. We have observed that in the purification process of A. simplex antigen by affinity chromatography, the majority of the proteins that produced cross-reactivity against A. suum and Toxocara canis were eliminated.
Rodero
Marta
M
Departamento de Parasitología, Facultad de Farmacia, Universidad Complutense, Madrid, Spain.
Cuéllar
Carmen
C
Fenoy
Soledad
S
del Aguila
Carmen
C
Chivato
Tomás
T
Mateos
José M
JM
Laguna
Rafael
R
D000818:Animals; D017129:Anisakiasis / Q000175:diagnosis*; D000947:Antigens, Helminth / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007075:Immunoglobulin M / Q000097:blood / Q000276:immunology; D007816:Larva Migrans, Visceral / Q000175:diagnosis*; D012680:Sensitivity and Specificity
D000947:Antigens, Helminth; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007073:Immunoglobulin E