Non-PCR-dependent detection of the factor V leiden mutation from genomic DNA using a homogeneous invader microtiter plate assay*

Ryan, Daniel; Nuccie, Bonnie; Arvan, Dean A.

doi:10.1016/s1084-8592(99)80037-x

Cited by 75 publications

(43 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some of these other defects are as easy to test for as factor V Leiden and can even be multiplexed in a single assay. [61][62][63] After factor V Leiden, the most common of the heritable thrombophilias are the prothrombin 20210A variant and hyperhomocysteinemia. The prothrombin variant is a single nucleotide change in the 3&cjs1227;-untranslated region of the gene that results in elevated circulating prothrombin levels through an unknown mechanism.…”

Section: Recommendationmentioning

confidence: 99%

American College of Medical Genetics Consensus Statement on Factor V Leiden Mutation Testing

Grody

Griffin

Taylor³

et al. 2001

Genetics in Medicine

156

118

View full text Add to dashboard Cite

SUMMARY OF ISSUES AND RECOMMENDATIONSIssue 1: Which methodology should be used: Factor V Leiden DNA testing or functional activated protein C (APC) resistance testing? Recommendation 1When appropriate clinical care requires testing for the factor V Leiden allele, either direct DNA-based genotyping or a factor V Leiden-specific functional assay is recommended. Patients who test positive by a functional assay should then be further studied with the DNA test for confirmation and to distinguish heterozygotes from homozygotes. Patients on heparin therapy or with known lupus anticoagulant should proceed directly to molecular testing if the modified functional assay is not used. When relatives of individuals known to have factor V Leiden are tested, the DNA method is recommended.Issue 2: Who should be tested? Recommendation 2Opinions and practices regarding factor V Leiden testing vary. Some physicians advocate testing of all patients with venous thrombosis except when active malignancy is present. Others exclude testing in patients over age 60 in the absence of a family history of thrombosis or a previous thrombotic event.There is growing consensus that testing should be performed in at least the following circumstances (these are the same general recommendations for testing for any thrombophilia): Age <50, any venous thrombosis. Venous thrombosis in unusual sites (such as hepatic, mesenteric, and cerebral veins). Recurrent venous thrombosis. Venous thrombosis and a strong family history of thrombotic disease. Venous thrombosis in pregnant women or women taking oral contraceptives. Relatives of individuals with venous thrombosis under age 50. Myocardial infarction in female smokers under age 50.Testing may also be considered in the following situations:Venous thrombosis, age >50, except when active malignancy is present. Relatives of individuals known to have factor V Leiden. Knowledge that they have factor V Leiden may influence management of pregnancy and may be a factor in decision-making regarding oral contraceptive use. Women with recurrent pregnancy loss or unexplained severe preeclampsia, placental abruption, intrauterine fetal growth retardation, or stillbirth. Knowledge of factor V Leiden carrier status may influence management of future pregnancies.Random screening of the general population for factor V Leiden is not recommended.Routine testing is not recommended for patients with a personal or family history of arterial thrombotic disorders (e.g., acute coronary syndromes or stroke) except for the special situation of myocardial infarction in young female smokers. Testing may be worthwhile for young patients (<50 years of age) who develop acute arterial thrombosis in the absence of other risk factors for atherosclerotic arterial occlusive disease. Neither prenatal testing nor routine newborn screening is recommended.Issue 3: Should testing be offered to individuals with environmental risk factors? Recommendation 3Factor V Leiden testing is recommended in women with venous thromboembolism during pregnancy or...

show abstract

Section: Recommendationmentioning

confidence: 99%

American College of Medical Genetics Consensus Statement on Factor V Leiden Mutation Testing

Grody

Griffin

Taylor³

et al. 2001

Genetics in Medicine

156

118

View full text Add to dashboard Cite

show abstract

“…The Invader ® assay could offer a simple diagnostic platform to detect mutations and single nucleotide polymorphisms (SNPs) with high specificity and high sensitivity, directly from unamplified genomic DNA in a homogeneous, isothermal, FRETbased format (32)(33)(34)(35). The Invader assay is a useful and powerful tool to identify HALP with CETP deficiency.…”

Section: Discussionmentioning

confidence: 99%

“…The Invader ® assay combines structure-specific cleavage enzymes and a universal fluorescent resonance energy transfer (FRET) system (32)(33)(34)(35). Primary probes and Invader oligonucleotide for each mutation were designed with Invader ® Creator software to have theoretic annealing temperatures of 63 Њ C and 77 Њ C, respectively, using a nearestneighbor algorithm on the basis of final probe and target concentrations.…”

Section: Detection Of Cetp Gene Mutations By Invader ® Assaymentioning

confidence: 99%

“…Primary probes and Invader oligonucleotide for each mutation were designed with Invader ® Creator software to have theoretic annealing temperatures of 63 Њ C and 77 Њ C, respectively, using a nearestneighbor algorithm on the basis of final probe and target concentrations. The primary probes and Invader oligonucleotides used to detect CETP gene mutations by Invader ® assay are shown by guest, on May 9, 2018 www.jlr.org…”

Section: Detection Of Cetp Gene Mutations By Invader ® Assaymentioning

confidence: 99%

See 1 more Smart Citation

Two novel missense mutations in the CETP gene in Japanese hyperalphalipoproteinemic subjects: High-throughput assay by Invader® assay

Nagano

Yamashita

Hirano

et al. 2002

Journal of Lipid Research

View full text Add to dashboard Cite

Cholesteryl ester transfer protein (CETP) deficiency is one of the most important and common causes of hyperalphalipoproteinemia (HALP) in the Japanese. CETP deficiency is thought to be a state of impaired reverse cholesterol transport, which may possibly lead to the development of atherosclerotic cardiovascular disease despite high HDL-cholesterol (HDL-C) levels. Thus, it is important to investigate whether HALP is caused by CETP deficiency. In the present study, we identified two novel missense mutations in the CETP gene among 196 subjects with a marked HALP (HDL-C ജ 2.59 mmol/l ‫؍‬ 100 mg/dl). The two missense mutations, L151P (CTC → CCC in exon 5) and R282C (CGC → TGC in exon 9), were found in compound heterozygous subjects with D442G mutation, whose plasma CETP levels were significantly lower when compared with those in D442G heterozygous subjects. In COS-7 cells expressing the wild type and mutant CETP, these two mutant CETP showed a marked reduction in the secretion of CETP protein into media (0% and 39% of wild type for L151P and R282C, respectively). These results suggested that two novel missense mutations cause the decreased secretion of CETP protein into circulation leading to HALP. By using the Invader ® assay for seven mutations, including two novel mutations of the CETP gene, we investigated their frequency among 466 unrelated subjects with HALP (HDL-C ജ 2.07 mmol/l ‫؍‬ 80 mg/dl). Two novel mutations were rare, but L151P mutation was found in unrelated subjects with a marked HALP.Furthermore, we demonstrated that CETP deficiency contributes to 61.7% and 31.4% of marked HALP and moderate HALP in the Japanese, respectively. Reverse cholesterol transport (RCT) is one of the major protective systems against atherosclerosis, in which small HDL or free apolipoprotein (apo)A-I removes cholesterol from the peripheral cells and delivers it to the liver (1, 2). We have investigated the molecular mechanisms of RCT by analyzing the pathophysiology of subjects with abnormal HDL metabolism and clarified the physiological significance of plasma cholesteryl ester transfer protein (CETP) and hepatic triglyceride lipase in human RCT (3,4). In this system, CETP is known to facilitate the transfer of cholesteryl ester (CE) from HDL to apoB-containing lipoproteins, and is one of the major determinants of plasma HDL-cholesterol (HDL-C) levels in humans (5)(6)(7)(8). This is true especially in the hypertriglyceridemic subjects (9). Furthermore, CETP affects not only plasma HDL-C levels but also the amount of small HDL particles, which may be more important for RCT (8,10).Genetic CETP deficiency is the most important and frequent cause of hyperalphalipoproteinemia (HALP) in the Japanese (11-21). Seven mutations have been demonstrated to cause HALP, including two common mutations: a G-to-A substitution at the 5 Ј splicing donor site of the intron 14 (Int14 ϩ 1 G → A) and a missense mutation of exon Abbreviations: BPI, bactericidal permeability increasing protein; CE, cholesteryl ester; CETP, cholesteryl ester tra...

show abstract

“…This assay can be applied to high-throughput genotyping (Lyamichev et al 1999;Mein et al 2000), but, although it does not require polymerase chain reaction (PCR) amplification of genomic DNA (Ryan et al 1999), it does require 100 ng of genomic DNA to assay a single SNP. Therefore, only 1000 SNPs can be genotyped from 100µg of genomic DNA (equivalent to a 5-to 10-ml sample of whole blood).…”

Section: Introductionmentioning

confidence: 99%

A high-throughput SNP typing system for genome-wide association studies

et al. 2001

View full text Add to dashboard Cite

One of the most difficult issues to be solved in genome-wide association studies is to reduce the amount of genomic DNA required for genotyping. Currently available technologies require too large a quantity of genomic DNA to genotype with hundreds or thousands of singlenucleotide polymorphisms (SNPs). To overcome this problem, we combined the Invader assay with multiplex polymerase chain reaction (PCR), carried out in the presence of antibody to Taq polymerase, as well as using a novel 384-well card system that can reduce the required reaction volume. We amplified 100 genomic DNA fragments, each containing one SNP, in a single tube, and analyzed each SNP with the Invader assay. This procedure correctly genotyped 98 of the 100 SNP loci examined in PCR-amplified samples from ten individuals; the genotypes were confirmed by direct sequencing. The reproducibility and universality of the method were confirmed with two additional sets of 100 SNPs. Because we used 40 ng of genomic DNA as a template for multiplex PCR, the amount needed to assay one SNP was only 0.4 ng; therefore, theoretically, more than 200,000 SNPs could be genotyped at once when 100 µg of genomic DNA is available. Our results indicate the feasibility of undertaking genome-wide association studies using blood samples of only 5-10 ml.

show abstract

Non-PCR-dependent detection of the factor V leiden mutation from genomic DNA using a homogeneous invader microtiter plate assay*

Cited by 75 publications

References 10 publications

American College of Medical Genetics Consensus Statement on Factor V Leiden Mutation Testing

American College of Medical Genetics Consensus Statement on Factor V Leiden Mutation Testing

Two novel missense mutations in the CETP gene in Japanese hyperalphalipoproteinemic subjects: High-throughput assay by Invader® assay

A high-throughput SNP typing system for genome-wide association studies

Contact Info

Product

Resources

About