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Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies
- Source :
- BMC Genetics, Vol 11, Iss 1, p 44 (2010), BMC Genetics
- Publication Year :
- 2010
- Publisher :
- Springer Science and Business Media LLC, 2010.
-
Abstract
- Background Mutations in TP53 are common events during carcinogenesis. In addition to gene mutations, several reports have focused on TP53 polymorphisms as risk factors for malignant disease. Many studies have highlighted that the status of the TP53 codon 72 polymorphism could influence cancer susceptibility. However, the results have been inconsistent and various methodological features can contribute to departures from Hardy-Weinberg equilibrium, a condition that may influence the disease risk estimates. The most widely accepted method of detecting genotyping error is to confirm genotypes by sequencing and/or via a separate method. Results We developed two new genotyping methods for TP53 codon 72 polymorphism detection: Denaturing High Performance Liquid Chromatography (DHPLC) and Dot Blot hybridization. These methods were compared with Restriction Fragment Length Polymorphism (RFLP) using two different restriction enzymes. We observed high agreement among all methodologies assayed. Dot-blot hybridization and DHPLC results were more highly concordant with each other than when either of these methods was compared with RFLP. Conclusions Although variations may occur, our results indicate that DHPLC and Dot Blot hybridization can be used as reliable screening methods for TP53 codon 72 polymorphism detection, especially in molecular epidemiologic studies, where high throughput methodologies are required.
- Subjects :
- Genotype
lcsh:QH426-470
Immunoblotting
Dot blot
Gene mutation
Biology
Methodology article
Nucleic Acid Denaturation
Denaturing high performance liquid chromatography
Genetics
Humans
Genetics(clinical)
Codon
Gene
Genotyping
Chromatography, High Pressure Liquid
Genetics (clinical)
Molecular Epidemiology
Molecular epidemiology
Nucleic Acid Hybridization
Genes, p53
lcsh:Genetics
Female
Restriction fragment length polymorphism
Polymorphism, Restriction Fragment Length
Subjects
Details
- ISSN :
- 14712156
- Volume :
- 11
- Database :
- OpenAIRE
- Journal :
- BMC Genetics
- Accession number :
- edsair.doi.dedup.....c8d931a47456a88f831823b82c8201b6
- Full Text :
- https://doi.org/10.1186/1471-2156-11-44