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

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 sclero sis complex (TSC) is an autosomal dominant familial tumor syndrome characte rized by the development of benign tumors (hamartomas) in multiple organs ( OMIM # 19110, # 191092). There is a high frequency of sporadic cases and si gnificant demand from patients and families for genetic testing information . Two TSC genes have been identified (TSC1 and TSC2) and together account f or all cases [1,2]. Here we report our methods for DHPLC analysis of the TS C1 gene and demonstrate the high sensitivity of this method in a blinded an alysis of 21 TSC patients with known TSC1 mutations. In this series, DHPLC detected 27/28 (96%) known TSC1 sequence variations. The only sequence vari ation not identified by DHPLC in this study is a mosaic case. (C) 2001 Else vier Science B.V. All rights reserved.