title: Pronto ThromboRisk--a novel primer-extension ELISA based assay for the detection of mutations associated with increased risk for thrombophilia.
abstract: The role of inherited thrombophilias in the pathogenesis of thrombosis, and their relation to thromboembolic events and pregnancy complications, has escalated the demand for molecular testing. We employed the new Pronto ThromboRisk kit (Pronto Diagnostics Ltd., Rehovot, Israel), which is based on a novel primer-extension ELISA assay, for the simultaneous detection of three genetic mutations. These are Factor V Leiden G1691A (R506Q), prothrombin G20210A, and methylenetetrahydrofolate reductase (MTHFR) C677T, which are known to have increased incidence in patients with thromboembolic events and pregnancy complications. We examined 284 randomly selected patient samples from the special coagulation hospital laboratory. The results using the new assay were compared to those obtained by routinely employed conventional molecular techniques. The ThromboRisk kit provided identical results with no false-negative results and with a high specificity of over 99% for all three mutations. We conclude that the Pronto ThromboRisk kit allows fast, precise, and reliable testing for the three genetic mutations. The assay is easy to perform and provides a useful tool for screening high risk populations, such as patients with a personal or family history of venous thromboembolism, women with pregnancy complications, or users of oral contraceptives.
authors.affiliation: The Hematology Institute, Tel Aviv Sourasky Medical Center, Israel.
authors.forename: Noga
authors.initials: N
authors.lastname: Carmi
authors.forename: Dana
authors.initials: D
authors.lastname: Cohen
authors.forename: Eti
authors.initials: E
authors.lastname: Zvang
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Naparstek
authors.forename: Varda
authors.initials: V
authors.lastname: Deutsch
mesh_terms: D017931:DNA Primers / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005165:Factor V / Q000235:genetics; D020022:Genetic Predisposition to Disease / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D006801:Humans; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques; D011516:Prothrombin / Q000235:genetics; D011933:Reagent Kits, Diagnostic; D019851:Thrombophilia / Q000150:complications / Q000175:diagnosis / Q000235:genetics*
chemical_list: D017931:DNA Primers; D011933:Reagent Kits, Diagnostic; C095381:factor V Leiden; D005165:Factor V; D011516:Prothrombin; D042965:Methylenetetrahydrofolate Reductase (NADPH2)