An improved methodology for the detection of the common mutation in the FGFR3 gene responsible for achondroplasia.

Homozygous achondroplasia is a neonatal lethal condition which can only be diagnosed in the first trimester of pregnancy by molecular analysis. The vast majority of patients with achondroplasia have a G-->A substitution at position 1138 of the fibroblast growth factor receptor (FGFR3) cDNA sequence, resulting in the substitution of an arginine for a glycine residue at position 380 of the FGFR3 protein. This mutation has typically been detected by SfcI digestion of amplified genomic DNA. We have demonstrated that the SfcI digestion protocol does not consistently distinguish between DNA samples heterozygous and homozygous for the G1138A substitution, and illustrates how the misdiagnosis of a homozygous affected fetus for one carrying only one copy of the G1138A mutation could occur. We report here an improved, simple nonradioactive technique which can reliably and consistently detect the presence of the G1138A mutation both in the heterozygous and homozygous state.