Arilla, M CIbarrola, IMartínez, AMonteseirín, JConde, JAsturias, J A2012-09-142012-09-142008-05-01Arilla MC, Ibarrola I, Martínez A, Monteseirín J, Conde J, Asturias JA. An antibody-based ELISA for quantification of Ani s 1, a major allergen from Anisakis simplex. Parasitology. 2008;135(6):735-401469-8161http://hdl.handle.net/10668/474Anisakis simplex is a nematode parasite that can infect humans who have eaten raw or undercooked seafood. Larvae invading the gastrointestinal mucosa excrete/secrete proteins that are implicated in the pathogenesis of anisakiasis and can induce IgE-mediated symptoms. Since Ani s 1 is a potent secreted allergen with important clinical relevance, its measurement could assess the quality of allergenic products used in diagnosis/immunotherapy of Anisakis allergy and track the presence of A. simplex parasites in fish foodstuffs. An antibody-based ELISA for quantification of Ani s 1 has been developed based on monoclonal antibody 4F2 as capture antibody and biotin-labelled polyclonal antibodies against Ani s 1 as detection reagent. The dose-response standard curves, obtained with natural and recombinant antigens, ranged from 4 to 2000 ng/ml and were identical and parallel to that of the A. simplex extract. The linear portion of the dose-response curve with nAni s 1 was between 15 and 250 ng/ml with inter-assay and intra-assays coefficients of variation less than 20% and 10%, respectively. The assay was specific since there was no cross-reaction with other extracts (except Ascaris extracts) and was highly sensitive (detection limit of 1·8 ng/ml), being able to detect Ani s 1 in fish extracts from codfish and monkfish.enAnisakis simplexAnisakis allergyQuantificationMonoclonal antibodyImmunoassayFood allergyRecombinant antigenParasite detectionAlergia a AnisakisCuantificaciónAnticuerpo monoclonalInmunoensayoAlergia a los alimentosAntígeno recombinanteDetección de parásitosMedical Subject Headings::Chemicals and Drugs::Biological Factors::Antigens::AllergensMedical Subject Headings::Diseases::Parasitic Diseases::Intestinal Diseases, Parasitic::AnisakiasisMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Blood Proteins::Immunoproteins::Immunoglobulins::Antibodies::Antibodies, HelminthMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Carrier Proteins::Calcium-Binding ProteinsMedical Subject Headings::Anatomy::Cells::Cells, Cultured::Cell Line::Cell Line, TumorMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Immunologic Techniques::Immunologic Tests::Immunoassay::Immunoenzyme Techniques::Enzyme-Linked Immunosorbent AssayMedical Subject Headings::Diseases::Animal Diseases::Fish DiseasesMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Helminth ProteinsMedical Subject Headings::Anatomy::Cells::Cells, Cultured::Clone Cells::HybridomasMedical Subject Headings::Organisms::Eukaryota::Animals::Animal Population Groups::Animals, Laboratory::Animals, Inbred Strains::Mice, Inbred Strains::Mice, Inbred BALB CMedical Subject Headings::Organisms::Eukaryota::Animals::Invertebrates::MolluscaMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Epidemiologic Methods::Statistics as Topic::Sensitivity and SpecificityMedical Subject Headings::Anatomy::Hemic and Immune Systems::Immune System::Lymphatic System::Lymphoid Tissue::SpleenMedical Subject Headings::Chemicals and Drugs::Biological Factors::Antigens::AllergensMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Blood Proteins::Immunoproteins::Immunoglobulins::Antibodies::Antibodies, HelminthMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Helminth ProteinsMedical Subject Headings::Organisms::Eukaryota::Animals::Invertebrates::Helminths::Nematoda::Secernentea::Ascaridida::Ascaridoidea::AnisakisMedical Subject Headings::Organisms::Eukaryota::Animals::Invertebrates::Arthropods::Crustacearesearch article18377696open access10.1017/S00311820080043320031-1820