Ion-pair reversed-phase denaturing HPLC-based biotechnology as a tool for genetic analysis

DHPLC (denaturing high-performance liquid chromatography) has established itself as one of the most powerful tools for DNA variation screening and allele discrimination for the characteristics of sensitivity, accuracy and efficiency that allow a rapid detection of SNPs, insertions and deletions. Moreover, it has the advantages of full automation and cost effectiveness. Aside from its well-known application in identifying mutations in human diseases, it has been also implemented in the quantitative measurement of gene expression and the analysis of single nucleotide extension products. In this paper, we aimed at summarizing the general properties and the main applications of DHPLC in the biomedical field as well as comparing this newly developed biotechnology with other conventional pre-screening methods for mutation scanning, such as single strand conformational polymorphisms (SSCP), conformation sensitive gel electrophoresis (CSGE) and two dimensional gene scanning (TDGS).