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Measurement of Factor V Activity in Human Plasma Using a Microplate Coagulation Assay
Published on: September 9, 2012
Development of a rapid DNA screening procedure for the Factor V Leiden mutation
1Division of Clinical Laboratory Sciences, Department of Clinical Chemistry, Queens Medical Centre, Nottingham.
Clinical Molecular Pathology
|December 1, 1996
Summary
A new DNA screening method using amplification refractory mutation system (ARMS) PCR rapidly and specifically detects the factor V Leiden mutation in whole blood. This test aids in investigating thrombovascular disease risk.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- The factor V Leiden mutation is a common genetic risk factor for thrombovascular disease.
- Accurate and rapid detection of this mutation is crucial for patient management.
Purpose of the Study:
- To develop a rapid, simple, and highly specific DNA screening procedure using ARMS PCR.
- To detect the factor V Leiden mutation in whole blood samples.
Main Methods:
- ARMS PCR amplification primers were designed with additional mismatches at the 3' end (-2 or -3 positions) to enhance specificity.
- These primers were used to detect both the normal Factor V gene and the Leiden mutation.
- A control set of primers was included for positive control.
Main Results:
- The developed ARMS assay successfully detected the factor V Leiden mutation in 0.5 μL of whole blood within three hours.
- Primers destabilized at position -3 allowed for lower annealing temperatures, increasing sensitivity and routine use.
- The assay demonstrated high specificity and rapidity.
Conclusions:
- This rapid and specific ARMS-based assay is a valuable tool for investigating factor V Leiden mutation.
- It aids in the evaluation of patients with or at risk of thrombovascular disease.
