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A relatively new method for the detection of single nucleotide polymorphisms is the use of denaturing highperformance liquid chromatography (DHPLC). DHPLC was used to analyse the transcobalamin II 776C&gt;G polymorphism in DNA from 159 healthy elderly. Furthermore, cobalamin, folate, homocysteine and holotranscobalamin II (holo-TC II) were measured. The allele frequency of the  G -allele was 17% with n = 55 harbouring the  CC  genotype, n = 77 being heterozygous and n = 27 showing the  GG  genotype. Holo-TC II concentrations were significantly decreased in patients harbouring the  GG  genotype. There was no effect on cobalamin, methylmalonyl-CoA, folate or homocysteine concentrations. A new G&gt;A variant at nucleotide position 810 in the  TC II  gene was detected by an altered peak pattern in the DHPLC and further elucidated by direct sequencing. The  TC II G810A  variant is a silent mutation without replacement of the corresponding amino acid (alanine) at position 270 in the TC II protein and was only found as a heterozygous genotype in a single patient.

clinical chemistry and laboratory medicine

Analysis of the Transcobalamin II 776C&gt;G (259P&gt;R) Single Nucleotide Polymorphism by Denaturing HPLC in Healthy Elderly: Associations with Cobalamin, Homocysteine and Holo-Transcobalamin II