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Analysis of the factor V Leiden mutation using the READIT Assay
R B Rhodes1, K Lewis, J Shultz
1University of Minnesota, Minneapolis, MN, USA.
Molecular Diagnosis : a Journal Devoted to the Understanding of Human Disease Through the Clinical Application of Molecular Biology
|March 21, 2001
Summary
The novel READIT Assay accurately detects the factor V Leiden mutation, showing 100% concordance in SNP genotyping. This luciferase-based method offers a straightforward and effective tool for genetic analysis.
Area of Science:
- Genetics
- Molecular Biology
Background:
- Single-nucleotide polymorphisms (SNPs) are crucial genetic markers.
- Existing methods for SNP detection in amplified DNA segments vary.
- The factor V Leiden mutation is a common genetic risk factor.
Purpose of the Study:
- To evaluate a novel SNP scoring tool, the READIT Assay, for analyzing the factor V Leiden mutation.
- To determine the accuracy and throughput of the READIT Assay.
Main Methods:
- A luciferase-based technique, READIT Technology, was employed.
- 510 residual human samples were analyzed for the factor V Leiden mutation.
- A blinded retrospective analysis was conducted.
Main Results:
- The READIT Assay demonstrated 100% concordance with existing genotype assignments.
- Significant separation ( > 6 SDs) was observed between wild-type and heterozygote populations.
- High separation ( > 9 SDs) was noted between heterozygote and homozygote populations.
Conclusions:
- The READIT Assay is an effective SNP scoring tool, achieving perfect concordance.
- The assay design is straightforward, utilizing unmodified oligonucleotides.
- Automated genotype assignments and statistical analysis are facilitated by the READIT Calculator.