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1 Deutsches Herzzentrum München and 1. Medizinische Klinik rechts der Isar, Technische Universität München, D-80636 München, Germany.
2 Technion—Israel Institute of Technology, The Bruce Rappaport Faculty of Medicine, Haifa, Israel.
aAddress correspondence to this author at: Deutsches Herzzentrum München, Experimentelle Kardiologie, Lazarettstrasse 36, D-80636 Munich, Germany. Fax 49-89-1218-3053; e-mail wkoch{at}dhm.mhn.de.
Background: A genetically defined molecular heterogeneity of haptoglobin, characterized by the major phenotypic forms Hp 1-1, Hp 2-1, and Hp 2-2, has been associated with distinct clinical manifestations. To enable the use of DNA samples for the study of this polymorphism, we established a haptoglobin genotyping method based on PCR.
Methods: Taking advantage of the selectivity of PCR, we amplified DNA segments specifically representing haptoglobin alleles Hp 1 and Hp 2 from genomic DNA. The products were analyzed by agarose gel electrophoresis. Haptoglobin phenotyping of plasma samples was performed by polyacrylamide gel electrophoresis and peroxidase staining.
Results: Exploiting the known size difference between Hp 1 and Hp 2, we amplified allele-specific DNA molecules with one pair of oligonucleotide primers. As an alternative, we used separate primer pairs to generate amplification products indicative of alleles Hp 1 and Hp 2. Because of the primer design, genotype determination was not compromised by sequence variations specifying haptoglobin allele subtypes S and F. For the same reason, the sequence similarity between the haptoglobin gene and the haptoglobin-related gene did not interfere with the accuracy of genotyping. Analysis with restriction enzymes demonstrated the authenticity of the allele-specific DNA products. Haptoglobin DNA genotyping and protein phenotyping, performed in parallel, yielded fully corresponding results. In a group of 249 individuals, the haptoglobin genotype distribution was as follows: 14.5% Hp 1-1, 48.2% Hp 2-1, and 37.3% Hp 2-2.
Conclusion: The new method can be used for genotyping of a common haptoglobin polymorphism.
The following articles in journals at HighWire Press have cited this article:
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  M. Soejima and Y. Koda TaqMan-Based Real-Time PCR for Genotyping Common Polymorphisms of Haptoglobin (HP1 and HP2) Clin. Chem., November 1, 2008; 54(11): 1908 - 1913. [Abstract] [Full Text] [PDF]
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T. Costacou, R. E. Ferrell, and T. J. Orchard Haptoglobin Genotype: A Determinant of Cardiovascular Complication Risk in Type 1 Diabetes Diabetes, June 1, 2008; 57(6): 1702 - 1706. [Abstract] [Full Text] [PDF]
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M. Papp, I. Foldi, E. Nemes, M. Udvardy, J. Harsfalvi, I. Altorjay, I. Mate, T. Dinya, C. Varvolgyi, Z. Barta, et al. Haptoglobin Polymorphism: A Novel Genetic Risk Factor for Celiac Disease Development and Its Clinical Manifestations Clin. Chem., April 1, 2008; 54(4): 697 - 704. [Abstract] [Full Text] [PDF]
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 U. Milman, S. Blum, C. Shapira, D. Aronson, R. Miller-Lotan, Y. Anbinder, J. Alshiek, L. Bennett, M. Kostenko, M. Landau, et al. Vitamin E Supplementation Reduces Cardiovascular Events in a Subgroup of Middle-Aged Individuals With Both Type 2 Diabetes Mellitus and the Haptoglobin 2-2 Genotype: A Prospective Double-Blinded Clinical Trial Arterioscler. Thromb. Vasc. Biol., February 1, 2008; 28(2): 341 - 347. [Abstract] [Full Text] [PDF]
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 M. Borsody, A. Burke, W. Coplin, R. Miller-Lotan, and A. Levy Haptoglobin and the development of cerebral artery vasospasm after subarachnoid hemorrhage Neurology, March 14, 2006; 66(5): 634 - 640. [Abstract] [Full Text] [PDF]
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M. Suleiman, D. Aronson, R. Asleh, M. R. Kapeliovich, A. Roguin, S. R. Meisel, M. Shochat, A. Sulieman, S. A. Reisner, W. Markiewicz, et al. Haptoglobin Polymorphism Predicts 30-Day Mortality and Heart Failure in Patients With Diabetes and Acute Myocardial Infarction Diabetes, September 1, 2005; 54(9): 2802 - 2806. [Abstract] [Full Text] [PDF]
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J. R. Delanghe and M. L. De Buyzere Development of an ELISA for the Determination of the Major Haptoglobin Phenotype: An Interesting Technical Development and Its Potential Consequences Clin. Chem., November 1, 2004; 50(11): 1972 - 1973. [Full Text] [PDF]
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N. S. Levy and A. P. Levy ELISA for Determination of the Haptoglobin Phenotype Clin. Chem., November 1, 2004; 50(11): 2148 - 2150. [Full Text] [PDF]
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 K U Park, J Song, and J Q Kim Haptoglobin genotypic distribution (including Hp0 allele) and associated serum haptoglobin concentrations in Koreans J. Clin. Pathol., October 1, 2004; 57(10): 1094 - 1095. [Abstract] [Full Text] [PDF]
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 C. Chiellini, F. Santini, A. Marsili, P. Berti, A. Bertacca, C. Pelosini, G. Scartabelli, E. Pardini, J. Lopez-Soriano, R. Centoni, et al. Serum Haptoglobin: A Novel Marker of Adiposity in Humans J. Clin. Endocrinol. Metab., June 1, 2004; 89(6): 2678 - 2683. [Abstract] [Full Text] [PDF]
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W. Koch, W. Latz, M. Eichinger, C. Gschwendner, B. Teige, A. Schomig, and A. Kastrati Haptoglobin Gene Subtyping by Restriction Enzyme Analysis Clin. Chem., November 1, 2003; 49(11): 1937 - 1940. [Full Text] [PDF]
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 A. Roguin, W. Koch, A. Kastrati, D. Aronson, A. Schomig, and A. P. Levy Haptoglobin Genotype Is Predictive of Major Adverse Cardiac Events in the 1-Year Period After Percutaneous Transluminal Coronary Angioplasty in Individuals With Diabetes Diabetes Care, September 1, 2003; 26(9): 2628 - 2631. [Abstract] [Full Text] [PDF]
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