高分辨率熔解曲线分析技术检测苯丙酮尿症患者的苯丙氨酸羟化酶基因突变

目的 应用高分辨率熔解曲线（HRM）分析技术检测苯丙酮尿症患者苯丙氨酸羟化酶基因突变.方法 利用PCR扩增13例苯丙酮尿症（PKU）患者的苯丙氨酸羟化酶（PAH）基因第6、7、11及12外显子,然后对PCR产物进行HRM分析,通过DNA测序对HRM结果进行验证.结果 在13例PKU患者的26个PAH等位基因中共检测出5种不同突变基因,总检出率为38.5％.常见的突变类型是R243Q和A434D.检测出两种多态性位点为Q232Q和V245V.HRM分析结果与测序结果完全一致.结论 HRM技术具有简单、快速、易操作、准确等优点,可用于PKU患者PAH基因突变筛查.

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

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.