Clinical Chemistry
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Clinical Chemistry 45: 41-46, 1999;
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(Clinical Chemistry. 1999;45:41-46.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Automated Detection of the Factor V Leiden Mutation Using the LCx Microparticle Enzyme Immunoassay

Mathilde Hunault1, Camile Marsh-Scott2, Cynthia Jou3, Ron Marshall3, Christi Scheffel3, Louis D. Fiore4, Kenneth A. Bauer1 and Michael J. Mitchell2,a

1 Hematology-Oncology Section, Brockton-West Roxbury Veterans Affairs Medical Center, Brockton, MA 02410.

2 Division of Laboratory Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215.

3 Abbott Laboratories, Molecular Diagnostics, Abbott Park, IL 60064.

4 Hematology-Oncology Section, Boston Veterans Affairs Medical Center, Boston, MA 02130.
a Address correspondence to this author at: Department of Hospital Laboratories, University of Massachusetts Medical Center, 50 Lake Avenue North, Worcester, MA 01655. Fax (508) 856-1537; e-mail Michael.Mitchell{at}banyan.ummed.edu.

The factor V Leiden mutation, a G->A transition at position 1691 in exon 10 of the gene that codes for factor V, produces an Arg506Gln substitution and is the most common genetic risk factor for venous thrombosis. We have developed a rapid, sensitive, and specific method to detect the factor V Leiden mutation in genomic DNA from whole blood by PCR amplification and microparticle enzyme immunoassay detection using the Abbott LCx instrument. We compared this automated method with the standard procedure using restriction endonuclease digestion of PCR products followed by gel electrophoresis in blinded experiments. In 130 patients (from Veterans Affairs medical centers) with deep venous thromboses, including 24 heterozygotes with the factor V Leiden mutation, there was complete agreement between the two methods. The assay was also able to distinguish heterozygotes from homozygotes. This method, which carries a low potential for cross-contamination of samples, should be a useful routine test for the factor V Leiden mutation in clinical laboratories with sufficient demand for molecular diagnostic assays using the LCx instrument.




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


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 CLIN APPL THROMB HEMOSTHome page
A.-M. Simundic, E. Topic, and M. Stefanovic
Detection of Factor V Leiden by PCR-SSCP using GMATM Precast Elchrom Scientific Gels
Clinical and Applied Thrombosis/Hemostasis, July 1, 2003; 9(3): 227 - 231.
[Abstract] [PDF]

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 Clin. Chem.Home page
J. G. Evans and C. Lee-Tataseo
Determination of the Factor V Leiden Single-Nucleotide Polymorphism in a Commercial Clinical Laboratory by Use of NanoChip Microelectronic Array Technology
Clin. Chem., September 1, 2002; 48(9): 1406 - 1411.
[Abstract] [Full Text] [PDF]


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