Automated detection of the G20210A prothrombin mutation using the LCx microparticle enzyme immunoassay

Clin Chim Acta. 2001 Dec;314(1-2):249-54. doi: 10.1016/s0009-8981(01)00655-6.

Abstract

Background: The prothrombin mutation, a G/A transition at position 20210 in the 3' untranslated region of the prothrombin gene, is associated with an increased risk of deep venous thrombosis and obstetrical complications. Several methods have been developed to detect the mutation; however, given the increased demand for this test in risk factor assessment, the development of simple and efficient screening methods has become necessary.

Methods: We have used a rapid, sensitive, and precise method developed by Abbott Laboratories to detect the prothrombin mutation. The method employs a polymerase chain reaction (PCR) amplification and the Abbot LCx microparticle enzyme immunoassay (MEIA) for detection. This method is able to detect and identify both homozygous and heterozygous genotypes.

Results: Two hundred ninety-six patients with a history of deep venous thrombosis, pulmonary embolism, preeclampsia, or cardiovascular disease and 163 control patients were included in this study. The prevalence of the mutation was 5.74% in the high-risk group and 3.06% in the control group. There was complete agreement between the results from the MEIA detection with those obtained using other detection methodologies, namely standard PCR and restriction fragment length polymorphism (RFLP) analysis.

Conclusions: The MEIA detection method of the prothrombin mutation represents a simple, fast, and reliable alternative to standard methods of detection and is well suited for use in routine clinical laboratories. The results of our study confirm others' studies showing a greater incidence of G20210A prothrombin gene mutation in patients with an increased risk of venous thrombosis and pulmonary embolism as well as patients with cardiovascular disease and pregnant women with preeclampsia. It reinforces the necessity of including the screening for prothrombin mutation in populations at risk.

Publication types

  • Clinical Trial
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions / genetics
  • Adolescent
  • Adult
  • Aged
  • Aged, 80 and over
  • DNA / genetics
  • DNA / isolation & purification
  • Electrophoresis, Polyacrylamide Gel
  • Female
  • Genetic Testing
  • Genotype
  • Humans
  • Immunoenzyme Techniques
  • Male
  • Microspheres
  • Middle Aged
  • Mutation
  • Prothrombin / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Risk Factors

Substances

  • 3' Untranslated Regions
  • Prothrombin
  • DNA