NONRADIOACTIVE SCREENING OF GLUCOKINASE MUTATIONS IN MATURITY-ONSET DIABETES OF THE YOUNG

DNA mutations were previously identified in the glucokinase gene in 56 % of French families affected with maturity onset diabetes of the youn g (MODY), an early onset autosomal dominant form of non-insulin-depend ent diabetes mellitus (NIDDM). Mutations were found on almost all exon s using the common radioactive single-strand conformation polymorphism (SSCP) technique. In this paper, we describe a non-isotopic SSCP meth od using the Pharmacia Biotech PhastSystem(TM) for the routine screeni ng of new mutations in diabetic patients or in offsprings of diabetic patients. The use of the PhastSystem allowed us to easily and reproduc ibly optimize the electrophoretic conditions for each exon. We demonst rate 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 reliab le method to improve the routine diagnosis of MODY in diabetic subject s or relatives and should be applicable to other genetic diseases.