title: Fully automated assay of glycohemoglobin with the Abbott IMx analyzer: novel approaches for separation and detection.
abstract: We describe a novel assay for measuring glycohemoglobin directly from anticoagulated whole blood with the Abbott IMx analyzer. The glycohemoglobin is labeled with a soluble polyanionic affinity reagent and the anionic complex is then captured with a cationic solid-phase matrix. Glycohemoglobin is quantified by measuring the quenching by heme of the static fluorescence from an added fluorophore. The assay is standardized to report both percent total glycohemoglobin (%GHb) and percent hemoglobin A1c (%HbA1c). Glucose, bilirubin, triglycerides, labile fraction, and hemoglobin variants do not interfere in the assay. Within- and between-run CVs are approximately 4-5%, with total CVs of approximately 6.5%. Highly significant linear correlations (r > 0.97) were obtained in comparison studies with two major assay methodologies. The time to obtain one result is approximately 10 min (including assay of a control), 56 min for 22 results. We describe the development, standardization, and validation of this new method.
authors.affiliation: Department of Thyroid, Metabolic, and Cardiovascular Diagnostics, Abbott Laboratories, Abbott Park, IL 60064.
authors.forename: D H
authors.initials: DH
authors.lastname: Wilson
authors.forename: J P
authors.initials: JP
authors.lastname: Bogacz
authors.forename: C M
authors.initials: CM
authors.lastname: Forsythe
authors.forename: P J
authors.initials: PJ
authors.lastname: Turk
authors.forename: T L
authors.initials: TL
authors.lastname: Lane
authors.forename: R C
authors.initials: RC
authors.lastname: Gates
authors.forename: D R
authors.initials: DR
authors.lastname: Brandt
mesh_terms: D001322:Autoanalysis / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D001663:Bilirubin / Q000097:blood; D001786:Blood Glucose / Q000378:metabolism; D005319:Fetal Hemoglobin / Q000032:analysis; D005454:Fluorescence Polarization; D006442:Glycated Hemoglobin / Q000032:analysis*; D006444:Hemoglobin C / Q000032:analysis; D006451:Hemoglobin, Sickle / Q000032:analysis; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D007124:Immunoenzyme Techniques; D011786:Quality Control; D012016:Reference Values; D012680:Sensitivity and Specificity; D014280:Triglycerides / Q000097:blood
chemical_list: D001663:Bilirubin; D001786:Blood Glucose; D005319:Fetal Hemoglobin; D006442:Glycated Hemoglobin; D006444:Hemoglobin C; D006451:Hemoglobin, Sickle; D014280:Triglycerides