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


     

Clinical Chemistry 45: 1898-1905, 1999;
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
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 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 Web of Science (41)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ørum, H.
Right arrow Articles by Borre, M. B.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ørum, H.
Right arrow Articles by Borre, M. B.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow Hemostasis and Thrombosis
(Clinical Chemistry. 1999;45:1898-1905.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Detection of the Factor V Leiden Mutation by Direct Allele-specific Hybridization of PCR Amplicons to Photoimmobilized Locked Nucleic Acids

Henrik Ørum1,a, Mogens H. Jakobsen2, Troels Koch1, Jens Vuust3 and Martin B. Borre1

1 PNA Diagnostics A/S, Rønnegade 2, DK-2100 Copenhagen, Denmark.

2 Exiqon A/S, Bygstubben 9, DK-2950 Vedbæk, Denmark.

3 Department of Clinical Biochemistry, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark.
a Author for correspondence. Fax 45-44-44-03-73; e-mail oerum{at}euroconnect.dk

Background: Individuals carrying the factor V Leiden mutation have been shown to have an increased risk of developing venous thromboembolism. Our aim was to develop an ELISA-like assay to detect the mutation in PCR-amplified genomic DNA using novel, high-affinity DNA analogs, termed locked nucleic acids (LNAs).

Methods: LNA octamer probes complementary to the factor V wild-type or mutated sequence were covalently attached to individual wells of a microtiter plate. Biotinylated factor V amplicons were added, and hybridization to the immobilized LNA probes was scored colorimetrically using a horseradish peroxidase-anti-biotin Fab conjugate and tetramethylbenzidine substrate.

Results: In a prospective study of 53 patients, the assay reproducibly scored both factor V homozygotes and heterozygotes with excellent sensitivity and specificity. All results were in complete agreement with the results obtained with the conventional PCR-restriction fragment length polymorphism technique.

Conclusions: The simplicity of the assay and its procedural relatedness to the widely used ELISA format should make it useful for routine factor V testing in the clinical laboratory.




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


Home page
 Nucleic Acids ResHome page
J. D. Levin, D. Fiala, M. F. Samala, J. D. Kahn, and R. J. Peterson
Position-dependent effects of locked nucleic acid (LNA) on DNA sequencing and PCR primers
Nucleic Acids Res., November 6, 2006; 34(20): e142 - e142.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
J. Rupp, W. Solbach, and J. Gieffers
Single-Nucleotide-Polymorphism-Specific PCR for Quantification and Discrimination of Chlamydia pneumoniae Genotypes by Use of a "Locked" Nucleic Acid.
Appl. Envir. Microbiol., May 1, 2006; 72(5): 3785 - 3787.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
Y. You, B. G. Moreira, M. A. Behlke, and R. Owczarzy
Design of LNA probes that improve mismatch discrimination.
Nucleic Acids Res., January 1, 2006; 34(8): e60 - e60.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. P. Johnson, L. M. Haupt, and L. R. Griffiths
Locked nucleic acid (LNA) single nucleotide polymorphism (SNP) genotype analysis and validation using real-time PCR
Nucleic Acids Res., March 26, 2004; 32(6): e55 - e55.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
D. A. Di Giusto and G. C. King
Strong positional preference in the interaction of LNA oligonucleotides with DNA polymerase and proofreading exonuclease activities: implications for genotyping assays
Nucleic Acids Res., February 18, 2004; 32(3): e32 - e32.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
D. A. Braasch, Y. Liu, and D. R. Corey
Antisense inhibition of gene expression in cells by oligonucleotides incorporating locked nucleic acids: effect of mRNA target sequence and chimera design
Nucleic Acids Res., December 1, 2002; 30(23): 5160 - 5167.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
N. Jacobsen, J. Bentzen, M. Meldgaard, M. H. Jakobsen, M. Fenger, S. Kauppinen, and J. Skouv
LNA-enhanced detection of single nucleotide polymorphisms in the apolipoprotein E
Nucleic Acids Res., October 1, 2002; 30(19): e100 - e100.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
A. Simeonov and T. T. Nikiforov
Single nucleotide polymorphism genotyping using short, fluorescently labeled locked nucleic acid (LNA) probes and fluorescence polarization detection
Nucleic Acids Res., September 1, 2002; 30(17): e91 - e91.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
N. Jacobsen, M. Fenger, J. Bentzen, S. L. Rasmussen, M. H. Jakobsen, J. Fenstholt, and J. Skouv
Genotyping of the Apolipoprotein B R3500Q Mutation Using Immobilized Locked Nucleic Acid Capture Probes
Clin. Chem., April 1, 2002; 48(4): 657 - 660.
[Full Text] [PDF]

Home page
 Clin. Chem.Home page
N. Zammatteo, L. Lockman, F. Brasseur, E. De Plaen, C. Lurquin, P.-E. Lobert, S. Hamels, T. Boon, and J. Remacle
DNA Microarray to Monitor the Expression of MAGE-A Genes
Clin. Chem., January 1, 2002; 48(1): 25 - 34.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1999 by the American Association for Clinical Chemistry.