title: Diagnostic ELISA for parietal cell autoantibody using tomato lectin-purified gastric H+/K(+)-ATPase (proton pump).
abstract: Circulating parietal cell autoantibodies, a useful diagnostic marker for autoimmune gastritis and pernicious anaemia, are currently routinely tested by serum immunofluorescence reactivity with frozen sections of rodent stomach. The major molecular targets of these parietal cell autoantibodies have recently been demonstrated to be the alpha- and the beta-subunits of the gastric H+/K(+)-ATPase (proton pump). We have demonstrated that tomato lectin binds specifically to the beta-subunit of the proton pump and concomittantly co-purifies the alpha-subunit. In the present study, we have exploited the latter observation for the development of a diagnostic ELISA for the detection of parietal cell autoantibodies and compared the performance of this assay with an ELISA using crude gastric membranes. The ELISAs were tested on 72 parietal cell autoantibody-positive sera, 72 parietal cell autoantibody-negative sera and 72 disease-control sera. The ELISA using lectin-purified canine proton pump was superior to that using crude canine gastric membranes in that it was about two-fold more sensitive (82% vs. 43%). With an assay sensitivity of 82% and a specificity of 90%, we propose that the ELISA using the lectin-purified proton pump is a rapid, simple, sensitive and specific diagnostic immunoassay for parietal cell autoantibodies.
authors.affiliation: Department of Pathology, Monash University Medical School, Monash University, Prahan, Victoria, Australia.
authors.forename: J S
authors.initials: JS
authors.lastname: Chuang
authors.forename: J M
authors.initials: JM
authors.lastname: Callaghan
authors.forename: P A
authors.initials: PA
authors.lastname: Gleeson
authors.forename: B H
authors.initials: BH
authors.lastname: Toh
mesh_terms: D001323:Autoantibodies / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005753:Gastric Mucosa / Q000201:enzymology; D006801:Humans; D037102:Lectins; D010295:Parietal Cells, Gastric / Q000201:enzymology / Q000276:immunology*; D037121:Plant Lectins; D000254:Sodium-Potassium-Exchanging ATPase / Q000276:immunology*
chemical_list: D001323:Autoantibodies; D037102:Lectins; D037121:Plant Lectins; C034981:tomato lectin; D000254:Sodium-Potassium-Exchanging ATPase