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Determination of Complement Factor H Functional Polymorphisms (V62I, Y402H, and E936D) using Sequence-Specific Primer PCR and Restriction Fragment Length Polymorphisms

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Abstract

Background: Complement factor H (CFH; HF) is an essential regulatory protein that plays a critical role in the homeostasis of the complement system in plasma. Several polymorphisms and mutations in the complement factor H gene (CFH; HF1) have been identified. These have revealed interesting associations with hemolytic-uremic syndrome and age-related macular degeneration.

Methods and Results: The aim of this study was to develop a rapid and reliable assay for determining genotypic variants of the CFH gene. Sequence-specific primer PCR and restriction fragment length polymorphism techniques were chosen for the analysis of CFH polymorphisms. The assays detected the following published single nucleotide polymorphisms of CFH in our Caucasian population (n = 271): rs800292, 257G→A (V62I); rs1061170, 1277T→C (Y402H); and rs1065489, 2881G→T (E936D). The allele frequencies (257G = 0.850, 1277T = 0.574, and 2881G = 0.839) that we obtained from a healthy Hungarian population were consistent with previously published results.

Conclusion: These analytical methods are simple, reliable, and rapid to perform, and are amenable to automation. Therefore, they could facilitate large-scale genotypic analyses of the CFH gene in various diseases, such as hemolytic-uremic syndrome, age-related macular degeneration, and cardiovascular diseases.

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Acknowledgments

This work was supported by the National Office for Research and Technology, and the National Research Fund (grant number T46837).

The authors have no conflicts of interest that are directly relevant to the content of this article.

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Authors and Affiliations

  1. Third Department of Internal Medicine, Faculty of Medicine, Szentágothai János Knowledge Center, Semmelweis University, Kúutvölgyi út 4, Budapest, H-1125, Hungary

    Adrienn Bíró, Zoltán Prohászka, George Füst &  Bernadett Blaskó

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  1. Adrienn Bíró
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  2. Zoltán Prohászka
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  3. George Füst
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  4. Bernadett Blaskó
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Correspondence to Bernadett Blaskó.

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Bíró, A., Prohászka, Z., Füst, G. et al. Determination of Complement Factor H Functional Polymorphisms (V62I, Y402H, and E936D) using Sequence-Specific Primer PCR and Restriction Fragment Length Polymorphisms. Mol Diag Ther 10, 303–310 (2006). https://doi.org/10.1007/BF03256205

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