HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit an electronic Letter to the Editor about this paper Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via ISI Web of Science (18) Citing Articles via Google Scholar Google Scholar Articles by Meyer, K. Articles by Ueland, P. M. Search for Related Content PubMed PubMed Citation Articles by Meyer, K. Articles by Ueland, P. M. Related Collections Molecular Diagnostics and Genetics Automation and Analytical Techniques

High-Level Multiplex Genotyping of Polymorphisms Involved in Folate or Homocysteine Metabolism by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

¹ LOCUS for Homocysteine and Related Vitamins, University of Bergen, Bergen, Norway.

^aAddress correspondence to this author at: LOCUS for Homocysteine and Related Vitamins, Department of Pharmacology, University of Bergen, Armauer Hansens Hus, N-5021 Bergen, Norway. Fax 47-55-974605; e-mail klaus.meyer{at}farm.uib.no.

Background: Increased plasma total homocysteine (tHcy), a risk factor for cardiovascular disease, is related to genetic, environmental, and nutritional factors, in particular folate status. Future large epidemiologic studies of the genetic basis of hyperhomocysteinemia will require high-throughput assays for polymorphisms of genes related to folate and Hcy metabolism.

Method: We developed a high-level multiplex genotyping method based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the detection of 12 polymorphisms in 8 genes involved in folate or Hcy metabolism. The assay includes methylenetetrahydrofolate reductase (MTHFR) 677C>T and 1298A>C, methionine synthase (MTR) 2756A>G, methionine synthase reductase (MTRR) 66A>G, cystathionine ß-synthase (CBS) 844ins68 and 699C>T, transcobalamin II (TCII) 776C>G and 67A>G, reduced folate carrier-1 (RFC1) 80G>A, paraoxonase-1 (PON1) 575A>G and 163T>A, and betaine homocysteine methyltransferase (BHMT) 742G>A.

Results: The failure rate of the assay was 1.7% and was attributable to unsuccessful DNA purification, nanoliter dispensing, and spectrum calibration. Most errors were related to identification of heterozygotes as homozygotes. The mean error rate was 0.26%, and error rates differed for the various single-nucleotide polymorphisms. Identification of CBS 844ins68 was carried out by a semiquantitative approach. The throughput of the MALDI-TOF MS assay was 1152 genotypes within 20 min.

Conclusions: This high-level multiplex method is able to genotype 12 polymorphisms involved in folate or Hcy metabolism. The method is rapid and reproducible and could facilitate large-scale studies of the genetic basis of hyperhomocysteinemia and associated pathologies.

The following articles in journals at HighWire Press have cited this article:

				S. E. Vollset, J. Igland, M. Jenab, A. Fredriksen, K. Meyer, S. Eussen, H. K. Gjessing, P. M. Ueland, G. Pera, N. Sala, et al. The Association of Gastric Cancer Risk with Plasma Folate, Cobalamin, and Methylenetetrahydrofolate Reductase Polymorphisms in the European Prospective Investigation into Cancer and Nutrition Cancer Epidemiol. Biomarkers Prev., November 1, 2007; 16(11): 2416 - 2424. [Abstract] [Full Text] [PDF]

				M. D Niculescu, K.-A. da Costa, L. M Fischer, and S. H Zeisel Lymphocyte gene expression in subjects fed a low-choline diet differs between those who develop organ dysfunction and those who do not Am. J. Clinical Nutrition, July 1, 2007; 86(1): 230 - 239. [Abstract] [Full Text] [PDF]

				A. Misra, J.-Y. Hong, and S. Kim Multiplex Genotyping of Cytochrome P450 Single-Nucleotide Polymorphisms by Use of MALDI-TOF Mass Spectrometry Clin. Chem., May 1, 2007; 53(5): 933 - 939. [Abstract] [Full Text] [PDF]

				J. K. Blievernicht, E. Schaeffeler, K. Klein, M. Eichelbaum, M. Schwab, and U. M. Zanger MALDI-TOF Mass Spectrometry for Multiplex Genotyping of CYP2B6 Single-Nucleotide Polymorphisms Clin. Chem., January 1, 2007; 53(1): 24 - 33. [Abstract] [Full Text] [PDF]

				K.-A. da Costa, O. G. Kozyreva, J. Song, J. A. Galanko, L. M. Fischer, and S. H. Zeisel Common genetic polymorphisms affect the human requirement for the nutrient choline FASEB J, July 1, 2006; 20(9): 1336 - 1344. [Abstract] [Full Text] [PDF]

				M. Kohlmeier, K.-A. da Costa, L. M. Fischer, and S. H. Zeisel Genetic variation of folate-mediated one-carbon transfer pathway predicts susceptibility to choline deficiency in humans PNAS, November 1, 2005; 102(44): 16025 - 16030. [Abstract] [Full Text] [PDF]

				N. Ghebranious, L. Ivacic, J. Mallum, and C. Dokken Detection of ApoE E2, E3 and E4 alleles using MALDI-TOF mass spectrometry and the homogeneous mass-extend technology Nucleic Acids Res., October 4, 2005; 33(17): e149 - e149. [Abstract] [Full Text] [PDF]

				M. Boniotto, M. H. Hazbon, W. J. Jordan, G. P. Lennon, J. Eskdale, D. Alland, and G. Gallagher Novel Hairpin-Shaped Primer Assay To Study the Association of the -44 Single-Nucleotide Polymorphism of the DEFB1 Gene with Early-Onset Periodontal Disease Clin. Vaccine Immunol., July 1, 2004; 11(4): 766 - 769. [Abstract] [Full Text]