Rapid, accurate genotyping of alcohol dehydrogenase-1B and aldehyde dehydrogenase-2 based on the use of denaturing HPLC
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Liang Li
Abstract
The genotypes of alcohol dehydrogenase-1B (ADH1B) and aldehyde dehydrogenase-2 (ALDH2) are related to alcohol dependence and some human disorders. Rapid, accurate genotyping methodologies for specific polymorphisms of these two genes are needed for molecular screening and testing of alcohol-related problems in populations. A polymerase chain reaction (PCR) composed of two separate amplicons was designed to generate sequences containing the polymorphic site of interest in the ADH1B and ALDH2 genes. The PCR amplicons for each sample were subjected to denaturing high-performance liquid chromatography (DHPLC), analysis performed under partially denaturing conditions as determined by profiling the mixture of a unique homozygous control and a tested sample amplicon. A total of 150 genomic DNA samples were tested to validate this assay by blind analysis. Direct DNA sequencing was performed on samples randomly selected from each of the genotype groups detected by DHPLC profiling. The results indicate 100% concordance between the sequencing analysis and the DHPLC detection. The method we present provides a reliable and fast genotyping procedure for molecular screening of alcohol-related problems.
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©2005 by Walter de Gruyter Berlin New York
Artikel in diesem Heft
- Contents Volume 43, 2005
- Author Index
- Subject Index
- ProteinChips: the essential tools for proteomic biomarker discovery and future clinical diagnostics
- Protein profiling as a diagnostic tool in clinical chemistry: a review
- Protein biochip systems for the clinical laboratory
- Automation of biochip array technology for quality results
- SELDI-TOF-MS proteomics of breast cancer
- Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development
- Separation of human serum proteins using the Beckman-Coulter PF2D™ system: analysis of ion exchange-based first dimension chromatography
- Rapid, accurate genotyping of alcohol dehydrogenase-1B and aldehyde dehydrogenase-2 based on the use of denaturing HPLC
- APOA1 polymorphisms are associated with variations in serum triglyceride concentrations in hypercholesterolemic individuals
- Simple PCR heteroduplex, SSCP mutation screening methods for the detection of novel catalase mutations in Hungarian patients with type 2 diabetes mellitus
- Glycogen phosphorylase BB in acute coronary syndromes
- Alteration in serum leptin correlates with alterations in serum N-telopeptide of collagen type I and serum osteocalcin during the progression of osteoporosis in ovariectomized rats
- Glycemic control in diabetes in three Danish counties
- Atorvastatin suppresses homocysteine formation in stimulated human peripheral blood mononuclear cells
- Buprenorphine detection in biological samples
- The effect of thyroid antibody positivity on reference intervals for thyroid stimulating hormone (TSH) and free thyroxine (FT4) in an aged population
- High-affinity antibodies in a new immunoassay for plasma tissue factor: reduction in apparent intra-individual variation
- Physiological matrix metalloproteinase concentrations in serum during childhood and adolescence, using Luminex® Multiplex technology
- Sensitive immunoassays for the autoantibodies reacting against citrullinated carboxy-terminal telopeptides of type I and type II collagens in patients with rheumatoid arthritis
- Acknowledgement
Artikel in diesem Heft
- Contents Volume 43, 2005
- Author Index
- Subject Index
- ProteinChips: the essential tools for proteomic biomarker discovery and future clinical diagnostics
- Protein profiling as a diagnostic tool in clinical chemistry: a review
- Protein biochip systems for the clinical laboratory
- Automation of biochip array technology for quality results
- SELDI-TOF-MS proteomics of breast cancer
- Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development
- Separation of human serum proteins using the Beckman-Coulter PF2D™ system: analysis of ion exchange-based first dimension chromatography
- Rapid, accurate genotyping of alcohol dehydrogenase-1B and aldehyde dehydrogenase-2 based on the use of denaturing HPLC
- APOA1 polymorphisms are associated with variations in serum triglyceride concentrations in hypercholesterolemic individuals
- Simple PCR heteroduplex, SSCP mutation screening methods for the detection of novel catalase mutations in Hungarian patients with type 2 diabetes mellitus
- Glycogen phosphorylase BB in acute coronary syndromes
- Alteration in serum leptin correlates with alterations in serum N-telopeptide of collagen type I and serum osteocalcin during the progression of osteoporosis in ovariectomized rats
- Glycemic control in diabetes in three Danish counties
- Atorvastatin suppresses homocysteine formation in stimulated human peripheral blood mononuclear cells
- Buprenorphine detection in biological samples
- The effect of thyroid antibody positivity on reference intervals for thyroid stimulating hormone (TSH) and free thyroxine (FT4) in an aged population
- High-affinity antibodies in a new immunoassay for plasma tissue factor: reduction in apparent intra-individual variation
- Physiological matrix metalloproteinase concentrations in serum during childhood and adolescence, using Luminex® Multiplex technology
- Sensitive immunoassays for the autoantibodies reacting against citrullinated carboxy-terminal telopeptides of type I and type II collagens in patients with rheumatoid arthritis
- Acknowledgement
