Crépin M et al. (2006) Evaluation of denaturing high performance liquid chromatography for the mutational analysis of the MEN1 gene. J Mol Endocrinol 36: 369–376

Currently, direct DNA sequencing is the method of choice for detecting mutations in MEN1 (the gene associated with multiple endocrine neoplasia type 1). As no mutational hotspots exist in MEN1, the entire gene must be sequenced, which is slow, labor intensive, and expensive. Denaturing high-performance liquid chromatography (DHPLC) is successfully used in the high-throughput, automated detection of mutations in other tumor-associated genes, and researchers in France have now confirmed that DHPLC can detect MEN1 mutations with 100% sensitivity, equivalent to the sensitivity of DNA sequencing. Crépin et al. retrospectively analyzed 160 samples from unrelated patients with MEN1 for whom sequence data were available, as well as positive and negative control samples. DHPLC results were obtained within a few hours.

MEN1 mutations reveal characteristic patterns on DHPLC, which differ from those generated by wild-type DNA, but confirmatory DNA sequencing is still required to identify specific mutations. Nonetheless, because DHPLC can reliably screen out samples containing only wild-type MEN1 and target potentially mutated DNA fragments, the authors estimate the overall cost of MEN1 genotypic diagnosis using DHPLC to be fivefold cheaper than diagnosis based on complete DNA sequencing. Neither DNA sequencing nor DHPLC can identify deletion or duplication of entire exons, however, detection of which requires quantitative analysis techniques.

DHPLC is based on the detection of DNA heteroduplexes formed in polymerase chain reaction products. The authors highlight that the specificity of DHPLC (98.6% in this study) depends on the purity of these products.