Multiplex ASA PCR for a Simultaneous Determination of Factor V Leiden Gene, G → A 20210 Prothrombin Gene and C → T677 MTHFR Gene Mutations

Hézard, Nathalie; Cornillet‐Lefèbvre, Pascale; Gillot, Lucile; Potron, G; Nguyen, Phu Cuong

doi:10.1055/s-0037-1615119

Cited by 55 publications

(19 citation statements)

References 5 publications

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“…DNA bands were separated by ethidium bromide-stained agarose (2% w/v) gel electrophoresis. In addition, both FV Leiden and PRT G20210A mutations were simultaneously analyzed by multiplex allele-specific amplification (ASA-PCR multiplex) as previously described [24]. Both assays gave 100% concordance rates.…”

Section: Genotype Analysismentioning

confidence: 99%

Association between adverse pregnancy outcomes and maternal factor V G1691A (Leiden) and prothrombin G20210A genotypes in women with a history of recurrent idiopathic miscarriages

et al. 2005

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Thrombophilia was implicated in the development of pregnancy complications, including recurrent idiopathic pregnancy loss, and is aggravated in women who are carriers of factor V G1691A (FV Leiden) and prothrombin (PRT) G20210A single-nucleotide polymorphisms (SNPs). Previous studies examined the role of FV-Leiden and PRT G20210A in recurrent pregnancy loss with conflicting results. Here we examined the prevalence of FV Leiden and PRT G20210A SNPs, in 200 women with 3 or more consecutive early (n = 87), late (n = 41), or early-late (n = 72) recurrent pregnancy losses, and 200 age-matched fertile parous control women. APC resistance (APCR) was detected functionally (measuring the activated clotting time triggered by activated factor X in presence of a fixed amount of purified APC), and FV-Leiden and PRT G20210A genotypes were assessed by PCR. The frequency of the mutant FV (0.1400 vs. 0.0276; P < 0.001) but not PRT 20210 (0.0100 vs. 0.0225; P = 0.159) allele was higher in patients than controls, respectively. APC resistance with factor V Leiden was seen in 27% of patients compared to 11.5% of controls, while APC resistance without factor V Leiden was seen in 12.5% of patients compared to 9.5% of controls. Regression analysis demonstrated that the significant predictors for early abortion was FV Leiden; those for late abortion were oral contraceptive, APCR, and FV Leiden; and predictors for early-late abortions were oral contraceptives, obesity, FV Leiden, and smoking. APC resistance and FV Leiden, as well as combination of both, are common thrombotic defects seen in women with idiopathic recurrent pregnancy loss, thus testing for these is recommended in women who have experienced recurrent miscarriages. Am.

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Section: Genotype Analysismentioning

confidence: 99%

Association between adverse pregnancy outcomes and maternal factor V G1691A (Leiden) and prothrombin G20210A genotypes in women with a history of recurrent idiopathic miscarriages

et al. 2005

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“…The presence of FVL was determined using an allele-specific (amplification refractory mutation system) polymerase chain reaction as previously described. 25,26 The HR2 was assessed using polymerase chain reaction and restriction fragment length polymorphism techniques adapted from the methods described by Bernardi et al 12 and Saiki et al 27 …”

Section: Genetic Analysismentioning

confidence: 99%

Activated Protein C Resistance Assay Detects Thrombotic Risk Factors Other Than Factor V Leiden

Graf¹,

Welsh

Qamar

et al. 2003

Am J Clin Pathol

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Activated protein C (APC) resistance is a common risk factor for venous thromboembolism (VTE) attributed to various mechanisms, including factor V Leiden (FVL) polymorphism. FVL is considered responsible for up to 95% of APC resistance; however, other factor V polymorphisms and elevated factor VIII levels also have been implicated. We assessed whether additional mechanisms contribute to APC resistance in a blinded case-control study of 65 subjects by measuring APC resistance using 3 methods: 2 activated partial thromboplastin time-based methods with and without dilution in factor V-deficient plasma and 1 Russell viper venom-based assay (RVV). Without factor V-deficient plasma, 24 subjects were APC resistant; with factor V-deficient plasma, the assay identified fewer APC-resistant subjects, as did RVV. All assays detected the 7 heterozygous FVL subjects. Thirteen subjects had factor VIII levels above 150% (1.50). After excluding subjects with FVL or elevated factor VIII levels, 4 subjects still had APC resistance. VTE risk trended higher for subjects with APC resistance in the absence of FVL. Measurement of APC resistance without dilution in factor V-deficient plasma is needed to assess for potentially important thrombotic risk factors other than FVL.

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“…The ASO hybridization approach has previously been applied to the detection of known point mutations underlying several genetic diseases. Detection of mutations in the CFTR gene (19) in Factor V, prothrombin, and MTHFR (20), as well as in mitochondrial DNA (21) was described.…”

Section: Resultsmentioning

confidence: 99%

Rapid Detection of CYP1A1, CYP2D6, and NAT Variants by Multiplex Polymerase Chain Reaction and Allele-Specific Oligonucleotide Assay

Labuda

Krajinović

Richer

et al. 1999

Analytical Biochemistry

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Multiplex ASA PCR for a Simultaneous Determination of Factor V Leiden Gene, G → A 20210 Prothrombin Gene and C → T677 MTHFR Gene Mutations

Cited by 55 publications

References 5 publications

Association between adverse pregnancy outcomes and maternal factor V G1691A (Leiden) and prothrombin G20210A genotypes in women with a history of recurrent idiopathic miscarriages

Association between adverse pregnancy outcomes and maternal factor V G1691A (Leiden) and prothrombin G20210A genotypes in women with a history of recurrent idiopathic miscarriages

Activated Protein C Resistance Assay Detects Thrombotic Risk Factors Other Than Factor V Leiden

Rapid Detection of CYP1A1, CYP2D6, and NAT Variants by Multiplex Polymerase Chain Reaction and Allele-Specific Oligonucleotide Assay

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