Application of natural and amplification created restriction sites for the diagnosis of PKU mutations

Abstract

PCR amplification, either conventional, or as site directed mutagenesls using primers with mismatched 3′-ends, followed by restriction endonuclease digestion, provides rapid, non-isotope assays of known mutations in the human phenylalanine hydroxylase gene. Such assays were shown to have the potential to detect all of the 18 presently reported phenyl-ketonuiia mutations. The practical applicability of this approach was demonstrated for eight mutations in Norwegian phenylketonuria patients, among them the most common ones.