A method to detect the G894T polymorphism of the NOS3 gene. Clinical validation in familial hypercholesterolemia
| Autor: | Leonardo de Oliveira Matsumoto, Rosario Dominguez Crespo Hirata, Luis A. Salazar, Mario Hiroyuki Hirata, Neusa Assumpta Forti, Jayme Diament, Kelson K. Yoshioka, Silvério Teixeira dos Santos, Sonia Q. Doi, Selma Andrea Cavalli, S D Giannini |
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| Rok vydání: | 2002 |
| Předmět: |
Adult
Male medicine.medical_specialty Adolescent Nitric Oxide Synthase Type III Clinical Biochemistry Familial hypercholesterolemia Polymorphism Single Nucleotide DNA sequencing law.invention Hyperlipoproteinemia Type II Exon Gene Frequency law medicine Humans Myocardial infarction Genetic Testing Deoxyribonucleases Type II Site-Specific Gene Polymerase chain reaction Polymerase Aged Genetics Aged 80 and over biology Biochemistry (medical) Cytogenetics General Medicine Middle Aged medicine.disease biology.protein Female Nitric Oxide Synthase Brazil |
| Zdroj: | Clinical chemistry and laboratory medicine. 40(5) |
| ISSN: | 1434-6621 |
| Popis: | 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 time-consuming. 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. |
| Databáze: | OpenAIRE |
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