title: Nonradioactive screening of glucokinase mutations in maturity onset diabetes of the young.
abstract: DNA mutations were previously identified in the glucokinase gene in 56% of French families affected with maturity onset diabetes of the young (MODY), an early onset autosomal dominant form of non-insulin-dependent diabetes mellitus (NIDDM). Mutations were found on almost all exons using the common radioactive single-strand conformation polymorphism (SSCP) technique. In this paper, we describe a non-isotopic SSCP method using the Pharmacia Biotech PhastSystem for the routine screening of new mutations in diabetic patients or in offsprings of diabetic patients. The use of the PhastSystem allowed us to easily and reproducibly optimize the electrophoretic conditions for each exon. We demonstrate the efficiency of this technique by identifying 8 mutations, 7 of which have never previously been detected, in patients referred to us for diagnostic purposes. It appears to be a sensible, easy and reliable method to improve the routine diagnosis of MODY in diabetic subjects or relatives and should be applicable to other genetic diseases.
authors.affiliation: Centre d'Etude du Polymorphisme Humain Fondation Jean Dausset-CEPH, Paris, France.
authors.forename: H
authors.initials: H
authors.lastname: Blanché
authors.forename: J
authors.initials: J
authors.lastname: Hager
authors.forename: F
authors.initials: F
authors.lastname: Sun
authors.forename: J
authors.initials: J
authors.lastname: Dausset
authors.forename: D
authors.initials: D
authors.lastname: Cohen
authors.forename: P
authors.initials: P
authors.lastname: Froguel
authors.forename: N
authors.initials: N
authors.lastname: Cohen
mesh_terms: D004252:DNA Mutational Analysis; D003924:Diabetes Mellitus, Type 2 / Q000201:enzymology / Q000235:genetics*; D005941:Glucokinase / Q000235:genetics*; D006801:Humans
chemical_list: D005941:Glucokinase