title: A simplification of the enzyme-linked immunospot technique. Increased sensitivity for cells secreting IgG antibodies to Haemophilus influenzae type b capsular polysaccharide.
abstract: A simplified enzyme-linked immunospot (ELISPOT) technique is described for the detection of cells secreting antibodies to tetanus toxoid (TT), diphtheria toxoid (DT) or Haemophilus influenzae type b capsular polysaccharide (PRP). By combining the cell suspension with the enzyme-linked secondary antibodies in one incubation, the second incubation and washing procedure could be omitted from the original technique. The simplified assay had the same sensitivity for anti-TT and anti-DT spot-forming cells as the ordinary ELISPOT assay. The IgG anti-PRP spots were, however, improved both in quality and in quantity (median: 40% more spots), while the detection of IgM and IgA anti-PRP spot-forming cells was the same in the two techniques. This simplified technique can probably also be used to save time in other antigen systems and should be considered when designing ELISPOT assays for the detection of polysaccharide-specific antibody-secreting cells.
authors: Laboratory of Medical Immunology, Medical Department TTA, Rigshospitalet, Copenhagen, Denmark.
authors: T
authors: T
authors: Barington
authors: S
authors: S
authors: Sparholt
authors: L
authors: L
authors: Juul
authors: C
authors: C
authors: Heilmann
mesh_terms: D000907:Antibodies, Bacterial / Q000032:analysis; D000942:Antigens, Bacterial / Q000276:immunology*; D016667:Bacterial Capsules / Q000276:immunology*; D004168:Diphtheria Toxoid / Q000276:immunology; D006193:Haemophilus influenzae / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007132:Immunoglobulin Isotypes / Q000032:analysis; D011135:Polysaccharides, Bacterial / Q000276:immunology*; D013745:Tetanus Toxoid / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D004168:Diphtheria Toxoid; D007132:Immunoglobulin Isotypes; D011135:Polysaccharides, Bacterial; D013745:Tetanus Toxoid