title: [Verification and Validation on Single Nucleotide Polymorphism Analysis of Alcohol Metabolism-Related Genes ADH1B and ALDH2, Using Dried-Saliva Samples].
abstract: We have developed a new method for unprocessed biological specimens as templates directly into the TaqMan assay. Saliva was needed to be put on a water-soluble paper and dried, because foreign substances, such as a filter paper, hinder fluorescence detection through the assay. Genotyping of alcohol metabolism-related genes ADH1B (rs1229984) and ALDH2 (rs671) polymorphisms was, subsequently, performed by TaqMan PCR assay using dried saliva in the present investigation. The optimized technique was tested on 114 samples of alcoholic patients. The PCR-RFLP methods with purified DNA from blood samples were employed for validation of the assay. Upon validation, complete concordance was observed between the two independent results. These results highlight the ability of TaqMan PCR assays using dried saliva on water-soluble paper in genotyping of ADH1B and ALDH2 genes. Our results showed a rapid, simple, reliable, and cost-effective method for SNP genotyping of mutations in ADH1B and ALDH2 genes. This will be very useful for large-scale association studies in various fields. [Original].
authors.forename: Shigenori
authors.initials: S
authors.lastname: Murata
authors.forename: Mariko
authors.initials: M
authors.lastname: Hayashida
authors.forename: Yuko
authors.initials: Y
authors.lastname: Ishiguro-Tanaka
authors.forename: Hiromi
authors.initials: H
authors.lastname: Imazeki
authors.forename: Emiko
authors.initials: E
authors.lastname: Hayashi
authors.forename: Akira
authors.initials: A
authors.lastname: Yokoyama
authors.forename: Kenji
authors.initials: K
authors.lastname: Kinoshita
mesh_terms: D000328:Adult; D000368:Aged; D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000444:Aldehyde Dehydrogenase / Q000235:genetics*; D000071396:Aldehyde Dehydrogenase, Mitochondrial; D000431:Ethanol / Q000378:metabolism*; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D010209:Paper; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D012463:Saliva; D012995:Solubility; D013048:Specimen Handling / Q000379:methods*; D014867:Water
chemical_list: D014867:Water; D000431:Ethanol; C489332:ADH1B protein, human; D000426:Alcohol Dehydrogenase; C096127:ALDH2 protein, human; D000444:Aldehyde Dehydrogenase; D000071396:Aldehyde Dehydrogenase, Mitochondrial