Robust nonradioactive oligonucleotide ligation assay to detect a common point mutation in the CYP2D6 gene causing abnormal drug metabolism
| Autor: | Ole Blaabjerg, Niels Erik Petersen, P Hyltoft-Petersen, Torben Hansen, A Iitiä, Mogens Hørder |
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| Rok vydání: | 1995 |
| Předmět: |
chemistry.chemical_classification
DNA ligase Oligonucleotide polymerase chain reaction time-resolved fluorescence assay Point mutation Biochemistry (medical) Clinical Biochemistry biotin- streptavidin CYP2D6 Gene Biology Molecular biology law.invention debrisoquine-sparteine polymorphism chemistry Biochemistry law DNA Mutational Analysis allele-specific ligation Ligation heritable disorders Polymerase chain reaction Drug metabolism |
| Zdroj: | Hansen, T S, Petersen, N E, Iitia, A, Blaabjerg, O, Hyltoft-Petersen, P & Horder, M 1995, ' Robust nonradioactive oligonucleotide ligation assay to detect a common point mutation in the CYP2D6 gene causing abnormal drug metabolism ', Clinical Chemistry, vol. 41, no. 3, pp. 413-418 . |
| ISSN: | 1530-8561 0009-9147 |
| DOI: | 10.1093/clinchem/41.3.413 |
| Popis: | A new nonradioactive oligonucleotide ligation assay for the detection of a common point mutation in the CYP2D6 gene is presented. The assay takes advantage of simultaneous time-resolved fluorescence measurements of lanthanide-labeled probes and the specificity of T4-DNA ligase in combination with the polymerase chain reaction. This strategy makes it possible to perform the assay using both the wild-type-specific and mutant-specific probes simultaneously, securing an internal control in each reaction. We show that the allele-specific ligation part of the assay can be performed with great accuracy over a wide range of temperatures, salt concentrations, and T4-DNA ligase concentrations. This eliminates the risk of false-positive or false-negative reactions due to variations in these factors. Because the assay is simple to perform, is very reliable, and can be partly automated, we conclude that it is well-suited for analysis in a routine laboratory. |
| Databáze: | OpenAIRE |
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