A RAPID PROTOCOL FOR CARDIAC TROPONIN-T GENE MUTATION DETECTION IN FAMILIAL HYPERTROPHIC CARDIOMYOPATHY

Mutations in the human cardiac troponin T gene (TNNT2) are associated with familial hypertrophic cardiomyopathy (FHC) linked to chromosome 1 q3 (CMH2). Mutation analyses of TNNT2 have been restricted to RNA base d screening methods because only the TNNT2 cDNA sequence was known. We characterized the genomic structure of 15 TNNT2 exons spliced into th e adult isoform. A protocol for rapid mutation detection based on dire ct sequencing of large PCR amplified genomic DNA fragments revealed a known TNNT2 mutation (Phe 110Ile) in one of 30 FHC probands. Three pol ymorphic short tandem repeat elements (D1S477, D1S2622, and D1S1723), useful for FHC pedigree analyses at CMH2, were shown to be physically tightly linked to TNNT2. (C) 1998 Wiley-Liss, Inc.