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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.

bmc genomic data

Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies