Denaturing high-performance liquid chromatography (DHPLC) is a highly sensitive, semi-automated method for identifying mutations in the TSC1 gene

https://doi.org/10.1016/S0165-022X(00)00149-4Get rights and content

Abstract

Sensitive and automated methods for the detection of DNA sequence variation are required for a wide variety of genetic studies. Diagnostic testing in human genetic disorders is one application of such methods. Tuberous sclerosis complex (TSC) is an autosomal dominant familial tumor syndrome characterized by the development of benign tumors (hamartomas) in multiple organs (OMIM # 19110, #191092). There is a high frequency of sporadic cases and significant demand from patients and families for genetic testing information. Two TSC genes have been identified (TSC1 and TSC2) and together account for all cases [1], [2]. Here we report our methods for DHPLC analysis of the TSC1 gene and demonstrate the high sensitivity of this method in a blinded analysis of 21 TSC patients with known TSC1 mutations. In this series, DHPLC detected 27/28 (96%) known TSC1 sequence variations. The only sequence variation not identified by DHPLC in this study is a mosaic case.

Section snippets

Web sites

  • http://zk.bwh.harvard.edu/projects/tsc/

  • http://insertion.stanford.edu/melt.html

  • http://www-genome.wi.mit.edu/cgi-bin/primer/primer3_www.cgi

  • http://www.ncbi.nlm.nih.gov/omim/

Acknowledgements

We wish to thank the TSC patients and families for contributing blood samples and/or financial support for this project. This work was supported by NIH grants CA71445 (SD) and NS 31535 (DK), and the National Tubersous Sclerosis Association.

References (17)

There are more references available in the full text version of this article.

Cited by (25)

  • Somatic Mosaicism and Neurological Diseases

    2016, Genomics, Circuits, and Pathways in Clinical Neuropsychiatry
  • DHPLC mutation analysis of phenylketonuria

    2003, Molecular Genetics and Metabolism
View all citing articles on Scopus
View full text