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Related Experiment Videos

An improved test to identify aPC-resistant factor V-Leiden

L Preda1, N Erba, S Figini

  • 1Immunohematology and Transfusion Service, Luigi Sacco Hospital, Milan, Italy.

Thrombosis Research
|June 15, 1997
PubMed
Summary

A new functional clotting assay effectively detects the factor V-Leiden mutation (R506Q) in patients on oral anticoagulants or with Lupus Anticoagulant inhibitors. This improved method provides reliable results where older assays failed.

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Area of Science:

  • Hematology
  • Clinical Chemistry
  • Thrombosis Research

Background:

  • The factor V-Leiden mutation (R506Q) is a common cause of inherited thrombophilia.
  • Traditional assays for activated protein C (aPC) resistance can yield unreliable results in patients with oral anticoagulants or Lupus Anticoagulant inhibitors.
  • Accurate detection of factor V-Leiden is crucial for managing thrombotic risk.

Purpose of the Study:

  • To develop and validate a modified functional clotting assay for detecting factor V-Leiden (R506Q) mutation.
  • To improve the reliability of factor V-Leiden detection in challenging patient populations.
  • To offer a robust diagnostic tool for thrombophilia screening.

Main Methods:

  • A modified prothrombin time (PT)-derived assay utilizing bovine thromboplastin and a 1:10 sample dilution was employed.

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  • The assay was evaluated for its performance in detecting heterozygous and homozygous factor V-Leiden mutations.
  • Assay automation and reproducibility were assessed on the ACL/3000 platform.

    • Main Results:

    • The modified assay demonstrated distinct clotting time prolongations for normal, heterozygous (10-18 seconds), and homozygous (<2 seconds) factor V-Leiden samples.
    • Reference normal plasma showed a prolongation exceeding 35 seconds under assay conditions.
    • The test proved to be easily automated, rapid, and exhibited good reproducibility.

    Conclusions:

    • The revised functional clotting assay provides a reliable method for detecting factor V-Leiden (R506Q) in patients, including those on oral anticoagulants or with Lupus Anticoagulant inhibitors.
    • This assay overcomes limitations of previous methods, offering improved diagnostic accuracy.
    • The assay's automation and reproducibility make it suitable for routine clinical laboratory use.