Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 43: 267-272, 1997;
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via ISI Web of Science (31)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ulvik, A.
Right arrow Articles by Ueland, P. M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ulvik, A.
Right arrow Articles by Ueland, P. M.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Endocrinology and Metabolism
(Clinical Chemistry. 1999;43:267-272.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

C677T mutation of methylenetetrahydrofolate reductase gene determined in blood or plasma by multiple-injection capillary electrophoresis and laser-induced fluorescence detection

Arve Ulvik1,a, Helga Refsum1, Leo A. J. Kluijtmans2 and Per Magne Ueland1

1 Department of Clinical Biology, Division of Pharmacology, University of Bergen, N-5021 Haukeland Hospital, Norway.

2 Department of Pediatrics, University Hospital Nijmegen, 6500 HB Nijmegen, The Netherlands.
a Author for correspondence. Fax +47-55-974605; e-mail arve.ulvik{at}ikb.uib.no

We constructed an assay to detect the common C677T mutation in the methylenetetrahydrofolate reductase gene. The mutation creates a Hinfl recognition site detected by restriction cleavage of a 198-bp fragment amplified in the polymerase chain reaction (PCR). Digested samples were subjected to capillary electrophoresis with laser-induced fluorescence detection (CE-LIF), with hydroxypropylmethylcellulose as the sieving matrix and SYBR Green I as the fluorescent dye. After amplification but before digestion, we added to the PCR mixture a fragment with the HinfI recognition site and a 15-bp truncation at the 3' end. Using this procedure, we could (a) verify completeness of digestion and monitor injection, (b) assign genotypes on the basis of pattern recognition, and (c) develop a multiple-injection mode with simultaneous separation of as many as eight samples. A seminested PCR protocol in combination with CE-LIF allowed genotyping of plasma/serum samples 20 years old.


Key Words: indexing terms: restriction enzyme analysis • genotyping • homocysteine • hyperhomocysteinemia • risk factors • premature atherosclerosis • cardiovascular disease • heritable disorders • folate deficiency




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


Home page
 Am. J. Clin. Nutr.Home page
R. Meleady, P. M Ueland, H. Blom, A. S Whitehead, H. Refsum, L. E Daly, S. E. Vollset, C. Donohue, B. Giesendorf, I. M Graham, et al.
Thermolabile methylenetetrahydrofolate reductase, homocysteine, and cardiovascular disease risk: the European Concerted Action Project
Am. J. Clinical Nutrition, January 1, 2003; 77(1): 63 - 70.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Clin. Nutr.Home page
H. Refsum, C. S Yajnik, M. Gadkari, J. Schneede, S. E Vollset, L. Orning, A. B Guttormsen, A. Joglekar, M. G Sayyad, A. Ulvik, et al.
Hyperhomocysteinemia and elevated methylmalonic acid indicate a high prevalence of cobalamin deficiency in Asian Indians
Am. J. Clinical Nutrition, August 1, 2001; 74(2): 233 - 241.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
A. Ulvik, J. Ren, H. Refsum, and P. M. Ueland
Simultaneous determination of methylenetetrahydrofolate reductase C677T and factor V G1691A genotypes by mutagenically separated PCR and multiple-injection capillary electrophoresis
Clin. Chem., February 1, 1998; 44(2): 264 - 269.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
R. Clarke, P. Woodhouse, A. Ulvik, C. Frost, P. Sherliker, H. Refsum, P. M. Ueland, and K.-T. Khaw
Variability and determinants of total homocysteine concentrations in plasma in an elderly population
Clin. Chem., January 1, 1998; 44(1): 102 - 107.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1997 by the American Association for Clinical Chemistry.