International reference standards: antibody standards for the indirect enzyme-linked immunosorbent assay.
Reference standards are used to calibrate similar assay systems against an international reference protocol and to provide a template for the preparation of secondary and/or working standards. Three reference standards are recommended for the indirect enzyme-linked immunosorbent assay: a strong positive standard, a weak positive standard and a negative serum standard. The negative standard should be derived from a single serum or from a serum pool which exhibits typical background activity in the reference protocol. The strong and weak positive standards should be derived from a single serum or from a serum pool which typifies the humoral response (antibody) to natural infection. Suitable candidates for the positive reference standards should exhibit dose/response curves in the mid-range of antibody activity. The strong and weak positive standards should each be prepared from a one-time dilution in the negative standard, to yield antibody activities which are defined by specific points on the linear portion of the dose/response curve. The strong positive standard should represent an antibody activity (absorbance value) midway between the upper and central points and the weak positive standard should represent an antibody activity midway between the central and lower points of the linear portion of the curve. Owing to inherent differences among assay systems, antibody activities should be expressed in relative rather than in absolute terms. It is recommended that the antibody activity of the strong positive standard should denote 100% positivity. The activities of the weak positive and negative standards should then be expressed as relative percentages. Every set of international reference standards should be accompanied by an information sheet which includes, among other things, a plot of the dose/response curve and an indication of the dilutions used to prepare the standards.
Wright
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PF
Canadian Food Inspection Agency, National Centre for Foreign Animal Disease, Winnipeg, Manitoba, Canada.
Tounkara
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K
Lelenta
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M
Jeggo
M H
MH
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D002003:Brucella abortus / Q000276:immunology*; D002007:Brucellosis, Bovine / Q000097:blood / Q000175:diagnosis* / Q000276:immunology; D002417:Cattle; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards / Q000662:veterinary*; D007391:International Cooperation; D008070:Lipopolysaccharides / Q000276:immunology; D011933:Reagent Kits, Diagnostic / Q000662:veterinary; D012015:Reference Standards; D015203:Reproducibility of Results
D000907:Antibodies, Bacterial; D008070:Lipopolysaccharides; D011933:Reagent Kits, Diagnostic