Abstract
Background: Haptoglobin (Hp) is an abundant serum protein which binds extracorpuscular hemoglobin (Hb). Two alleles exist in humans for the Hp gene, denoted 1 and 2. Diabetic individuals with the Hp 2-2 genotype are at increased risk of developing vascular complications including heart attack, stroke, and kidney disease. Recent evidence shows that treatment with vitamin E can reduce the risk of diabetic vascular complications by as much as 50% in Hp 2-2 individuals. We sought to develop a rapid and accurate test for Hp phenotype (which is 100% concordant with the three major Hp genotypes) to facilitate widespread diagnostic testing as well as prospective clinical trials.
Methods: A monoclonal antibody raised against human Hp was shown to distinguish between the three Hp phenotypes in an enzyme linked immunosorbent assay (ELISA). Hp phenotypes obtained in over 8000 patient samples using this ELISA method were compared with those obtained by polyacrylamide gel electrophoresis or the TaqMan PCR method.
Results: Our analysis showed that the sensitivity and specificity of the ELISA test for Hp 2-2 phenotype is 99.0% and 98.1%, respectively. The positive predictive value and the negative predictive value for Hp 2-2 phenotype is 97.5% and 99.3%, respectively. Similar results were obtained for Hp 2-1 and Hp 1-1 phenotypes. In addition, the ELISA was determined to be more sensitive and specific than the TaqMan method.
Conclusions: The Hp ELISA represents a user-friendly, rapid and highly accurate diagnostic tool for determining Hp phenotypes. This test will greatly facilitate the typing of thousands of samples in ongoing clinical studies.
This work was supported by NIH RO1DK085226-03 to A.P.L. and NIH 1RO1DK-077510 to A.D.P. We thank Michael Mauer for providing access to samples from the RAR study. The Diabetes-Dementia study was supported by NIH RO1 AG034087 to Michal Beeri. The CACTI study was supported by NHLBI RO1 HL61753 and HL079611, an American Diabetes Association Junior Faculty Award 1-10-JF-50 (J.S.B.), and the Diabetes Endocrinology Research Center Clinical Investigation Core P30 DK57516. The CACTI study was performed at the Clinical Translational Research Center at the University of Colorado Denver supported by NIH MO1 RR000051. A complete list of participants in the DCCT/EDIC research group can be found in New England Journal of Medicine 2011;365:2366–76. Contributors of free or discounted supplies and/or equipment: industry contributors have had no role in the DCCT/EDIC study but have provided free or discounted supplies or equipment to support participants’ adherence to the study: Abbott Diabetes Care (Alameda, CA), Animas (Westchester, PA), Bayer Diabetes Care (North America Headquarters, Tarrytown NY), Becton Dickinson (Franklin Lakes, NJ), CanAm (Atlanta, GA), Eli Lilly (Indianapolis, IN), Lifescan (Milpitas, CA), Medtronic Diabetes Minneapolis, MI), Omron (Shelton, CT), OmniPod® Insulin Management System (Bedford, MA), Roche Diabetes Care (Indianapolis, IN), and Sanofi-Aventis (Bridgewater NJ). Acknowledgement for the DCCT/EDIC has been supported by U01 Cooperative Agreement grants (1983–1993) and contracts (1994 to present) with the Division of Diabetes Endocrinology and Metabolic Diseases of the National Institute of Diabetes and Digestive and Kidney Disease, and through support by the National Eye Institute and by Genentech through a Cooperative Research and Development Agreement with the National Institute of Diabetes and Digestive and Kidney Diseases, the National Institute of Neurologic Disorders and Stroke, the Genetic Clinical Research Centers Program (1993–2007), and Clinical Translational Science Center Program (2006 to present), Bethesda, MD, USA.
Conflict of interest statement
Authors’ conflict of interest disclosure: APL and the Rappaport Institute are the owners of a patent for haptoglobin phenotype testing for the determination of vascular complications in diabetic individuals. Research support played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
Research funding: See Acknowledgments.
Employment or leadership: None declared.
Honorarium: None declared.
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