title: The development of rapid and accurate screening test for RET hotspot somatic and germline mutations in MEN2 syndromes.
abstract: Medullary thyroid carcinoma (MTC) is a rare endocrine malignancy with distinctive features separating it from other thyroid cancers. Cancer may be sporadic or occur as a consequence of the hereditary syndrome called multiple endocrine neoplasia type 2 (MEN2) with three distinct phenotypes in MEN2A, MEN2B and FMTC. Each variant of MEN2 results from different RET gene mutations, with a good genotype-phenotype correlation. The goal of the study was to develop a fast and accurate screening method for a reliable detection of hot-spot RET germline and sporadic tumor mutations. From a cohort of 191 patients with MTC and their relatives, 38 tested positive and 31 tested negative for a germline or somatic tumor RET mutation were selected. A positive HRM mutation pattern was detected in all mutation-positive patients and altogether the method was able to clearly differentiate between twenty different genotypes. A novel germline variant p.Ala639Thr was detected in MTC patient, which was determined to be likely benign. Analytical specificity was determined to be 98.6% and a sensitivity threshold was determined to be 30%. The fast and accurate HRM method reduces the turnaround time providing fast and important information, especially when targeted anti-tyrosine kinase therapy on tumor samples is considered. Overall, we developed a high-throughput, accurate and cost-effective approach for the detection of RET germline and sporadic tumor mutations.
authors: Department of Molecular Genetics, Institute of Pathology, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, Slovenia. Electronic address: andrej.zupan@mf.uni-lj.si.
authors: Andrej
authors: A
authors: Zupan
authors: Department of Molecular Genetics, Institute of Pathology, Faculty of Medicine, University of Ljubljana, Zaloška cesta 4, 1000 Ljubljana, Slovenia. Electronic address: damjan.glavac@mf.uni-lj.si.
authors: Damjan
authors: D
authors: Glavač
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods; D005091:Exons; D056726:Genetic Association Studies / Q000379:methods; D005820:Genetic Testing / Q000379:methods*; D005838:Genotype; D018095:Germ-Line Mutation / Q000235:genetics*; D006801:Humans; D018813:Multiple Endocrine Neoplasia Type 2a / Q000235:genetics*; D018814:Multiple Endocrine Neoplasia Type 2b / Q000235:genetics*; D010641:Phenotype; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics*
chemical_list: C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human
keywords: High resolution melt analysis; MEN2A; MEN2B; Multiple endocrine neoplasia; RET proto-oncogene