title: The minisequencing method: a simple strategy for genetic screening of MEN 2 families.
abstract: BACKGROUND
  Multiple endocrine neoplasia type 2 is an autosomal dominant disorder. MEN 2A is characterized by medullary thyroid carcinoma, pheochromocytoma and hyperparathyroidism; MEN 2B by medullary thyroid carcinoma, pheochromocytoma and characteristic stigmata. Activating germline mutations of the RET proto oncogene are responsible for this hereditary syndrome. Codon 634 mutations are the most common mutations occurring in MEN 2A families whereas a specific mutation at codon 918 is observed in the great majority of MEN 2B families. Analysis of these codons will provide a final diagnosis in the great majority of affected families making unnecessary further studies. To specifically study the codons 634 and 918 we used a minisequencing method as an alternative method to complete sequencing.
  
  
  RESULTS
  Using this mutation detection method we were able to reproduce in all cases, representative of 7 families, the information previously obtained by direct sequencing of PCR products. Depending on the number of primers used in the minisequencing reaction, we were able to interrogate either only one nucleotide of the target codon or the three nucleotides simultaneously.
  
  
  CONCLUSIONS
  This technique appears as a simple, rapid and efficient method for genetic screening of MEN 2 families. It can be utilized to seek for unknown mutations at specific codons or to screen for previously identified mutations and is therefore of interest to study index cases or individuals at risk. Results suggest that complete sequencing is unnecessary.
authors.affiliation: Centro de Investigação de Patobiologia Molecular, Instituto Português de Oncologia FG / Lisboa, Portugal. mjbugalho@ipolisboa.min-saude.pt
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Bugalho
authors.forename: Rita
authors.initials: R
authors.lastname: Domingues
authors.forename: Luís
authors.initials: L
authors.lastname: Sobrinho
mesh_terms: D003062:Codon; D004252:DNA Mutational Analysis / Q000379:methods*; D029721:Drosophila Proteins; D019075:Electrophoresis, Capillary; D005192:Family Health; D005820:Genetic Testing / Q000379:methods*; D018095:Germ-Line Mutation; D006801:Humans; D018813:Multiple Endocrine Neoplasia Type 2a / Q000175:diagnosis*; D018814:Multiple Endocrine Neoplasia Type 2b / Q000175:diagnosis*; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins / Q000235:genetics; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics
chemical_list: D003062:Codon; D029721:Drosophila Proteins; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D011518:Proto-Oncogene Proteins; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases; C416649:Ret protein, Drosophila