title: Efficient mutation detection in MEN1 gene using a combination of single-strand conformation polymorphism (MDGA) and heteroduplex analysis.
abstract: For facilitated genotypic analysis of multiple endocrine neoplasia type 1 (MEN1), a familial syndrome associated with tumors of the parathyroid and neuroendocrine tissues, we developed two screening methods, heteroduplex mutation assay (HMA) and mutation detection gel analysis (MDGA), both based on electrophoretic discrimination of polymerase chain reaction (PCR) products, to detect the mutations. Forty-three genomic DNA samples were used for the evaluation of these techniques. The whole coding region of MEN1 was PCR-amplified with fluorescent primers and then denatured/renatured before electrophoresis on an automated sequencer. 100% of the mutations were detected, subsequently confirmed and identified by sequencing. "Negative" samples were used to evaluate the specificity and reproducibility of the two techniques. The combination of the two methods allows high throughput cost-effective mutation screening which is less laborious than systematic sequencing of the whole coding region of MEN1. Together, these methods provide an efficient screen for MEN1 mutations.
authors.affiliation: Secteur Oncologie Moléculaire Huriez-USN, Laboratoire de Biochimie et Biologie Moléculaire, CHRU, Lille, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Crépin
authors.forename: Fabienne
authors.initials: F
authors.lastname: Escande
authors.forename: Pascal
authors.initials: P
authors.lastname: Pigny
authors.forename: Marie-P
authors.initials: MP
authors.lastname: Buisine
authors.forename: Alain
authors.initials: A
authors.lastname: Calender
authors.forename: Nicole
authors.initials: N
authors.lastname: Porchet
authors.forename: Marie-Françoise
authors.initials: MF
authors.lastname: Odou
mesh_terms: D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods* / Q000706:statistics & numerical data; D017931:DNA Primers / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D016147:Genes, Tumor Suppressor; D020180:Heteroduplex Analysis / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D018761:Multiple Endocrine Neoplasia Type 1 / Q000235:genetics; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011518:Proto-Oncogene Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; C105373:MEN1 protein, human; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins