A Method to Detect the G894T Polymorphism of the NOS3 Gene. Clinical Validation in Familial Hypercholesterolemia
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Rosario D.C. Hirata
Abstract
An endothelial nitric oxide synthase gene (NOS3) polymorphism in exon 7 (G894T), resulting in Glu298Asp substitution at protein level, has been associated with myocardial infarction, hypertension and coronary atherosclerosis in some populations. This polymorphism is usually identified by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). However, the procedures described to date do not eliminate the possibility of misclassification and either require confirmation by DNA sequencing or are timeconsuming. In this study, a PCR-RFLP procedure to detect the G894T polymorphism at the NOS3 was optimized by the introduction of a constitutive cleavage site in the amplification product. This cleavage site provides an internal control for enzymatic activity to avoid mistyping. The method was validated by the study of 35 white unrelated individuals with familial hypercholesterolemia and 70 controls. The frequency of the variant allele (T) was similar between both groups (27% vs. 22%, NS), and comparable to the frequency found in other white populations. However, future studies are necessary to confirm these data. In summary, the optimized procedure for detection of the G894T NOS3 polymorphism is rapid, simple, and does not require confirmatory tests. Using this method, we found no association between this polymorphism and familial hypercholesterolemia.
Copyright © 2002 by Walter de Gruyter GmbH & Co. KG
Artikel in diesem Heft
- XV Latin American Congress of Clinical Biochemistry, Florianópolis, Brazil, 1–5 July 2001
- A Method to Detect the G894T Polymorphism of the NOS3 Gene. Clinical Validation in Familial Hypercholesterolemia
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- IFCC News: May/June 2002
- Meetings and Awards
Artikel in diesem Heft
- XV Latin American Congress of Clinical Biochemistry, Florianópolis, Brazil, 1–5 July 2001
- A Method to Detect the G894T Polymorphism of the NOS3 Gene. Clinical Validation in Familial Hypercholesterolemia
- Increasing the Sensitivity of Single-Strand Conformation Polymorphism Analysis of the LDLR Gene Mutations in Brazilian Patients with Familial Hypercholesterolemia
- Hormone Replacement Therapy in Postmenopausal Women and Its Effects on Plasma Lipid Levels
- The Effect of N-Acetylcysteine Supplementation upon Viral Load, CD4, CD8, Total Lymphocyte Count and Hematocrit in Individuals Undergoing Antiretroviral Treatment
- An Evaluation of Antiretroviral Therapy Associated with α-Tocopherol Supplementation in HIV-infected Patients
- Effects of Re-education in Eating Habits and Physical Activity on the Lipid Profile of Obese Teenagers
- Myeloperoxidase-mediated Protein Oxidation: Its Possible Biological Functions
- Novel Haptoglobin Insertion/Deletion Polymorphism Is Associated with the Lipid Profile and C-Reactive Protein (CRP) Concentration
- Apolipoprotein E2/E2 Genotype in Combination with Mutations in the LDL Receptor Gene Causes Type III Hyperlipoproteinemia
- High-Sensitivity Human Thyroglobulin (hTG) Immunoradiometric Assay in the Follow-up of Patients with Differentiated Thyroid Cancer
- Tissue Transglutaminase-Serology Markers for Coeliac Disease
- Serum Adenosine Deaminase and Cytidine Deaminase Activities in Patients with Systemic Lupus Erythematosus
- Effects of Oral N-Acetylcysteine on Plasma Homocysteine and Whole Blood Glutathione Levels in Healthy, Non-pregnant Women
- Evaluating Sequential Values Using Time-adjusted Biological Variation
- High-dose Methylprednisolone Therapy in Multiple Sclerosis Increases Serum Uric Acid Levels
- Evaluation of Accuracy and Uncertainty of ELISA Assays for the Determination of Interleukin-4, Interleukin-5, Interferon-γ and Tumor Necrosis Factor-α
- Development of Immunoturbidimetric Assays for Fourteen Human Serum Proteins on the Hitachi 912™
- Multicenter Evaluation of a Fully Mechanized Soluble Transferrin Receptor Assay on the Hitachi and Cobas Integra Analyzers. The Determination of Reference Ranges
- IFCC News: May/June 2002
- Meetings and Awards
