Detection of phenylalanine hydroxylase gene mutations in patients wth phenylketonuria by high-resolution melting curve analysis

Abstract

Objective To identify patients with phenylketonuria (PKU) based on the detection of phenylalanine hydroxylase (PAH) gene mutations using high-resolution melting (HRM) analysis.Methods Exons-6,-7,-11 and-12 of PAH gene in 13 patients with PKU were amplified by polymerase chain reaction.This was followed by HRM analysis on the products of polymerase chain reaction according to gene sequencing.Results Of 26 alleles of PAH genes derived from 13 patients with PKU,5 mutants,including R243Q and A434D,were identified,thus yielding a positivity of 38.5％.Two polymorphism sites (Q232Q and V245V) were also identified,suggesting that our results were consistent with those of DNA sequencing.Conclusion HRM can be employed for screening of gene mutation in patients with PKU owing to the merits of being simple,rapid-responded,convenient and accurate. Key words: Phenylketonurias ; Phenylalanine hydroxylase ; Point mutation ; High-resolution melting curve