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目的建立一种简便、准确和快速的筛查苯丙酮尿症（ＰＫＵ）突变基因的方法。方法应用生物素渗入的聚合酶链反应扩增中国人ＰＫＵ患者常见突变位点：Ｙ２０４Ｃ（ｅｘｏｎ６，Ｅ６）、Ｒ２４３Ｑ（Ｅ７）、Ｙ３５６Ｘ（Ｅ１１）和Ｒ４１３Ｐ（Ｅ１２）所在的４个外显子区域，扩增标记产物再与固定于同一张膜上的等位基因特异性寡核苷酸探针进行逆相点杂交（ＲＤＢ），用碱性磷酸酶显色法检测杂交信号以确定突变类型。结果建立了检测以上突变位点的非同位素逆相点杂交方法，并对５例就诊的ＰＫＵ患者进行了ＲＤＢ检测，查明３例携带Ｒ２４３Ｑ突变，其中１例还带有Ｙ３５６Ｘ突变，用单链构象多态性方法验证了上述结果。结论该方法适用于对中国人ＰＫＵ患者进行常见突变位点的快速筛查
Objective To establish a simple, accurate and rapid screening method for gene mutations in phenylketonuria (PKU). Methods The common mutation sites of PKU patients in China were amplified by polymerase chain reaction with biotin. Four exon6 (exon6, E6), R243Q (E7), Y356X (E11) and R413P The amplified product was then reverse dot blot (RDB) with allele-specific oligonucleotide probes immobilized on the same membrane. The hybridization signal was detected by alkaline phosphatase assay to determine the type of mutation. Results A non-isotopic reverse-phase hybridization method was established to detect the above mutation sites. RDB detection was performed in 5 patients with PKU. Three cases of R243Q mutation were found, of which 1 case also had Y356X mutation, Conformational polymorphism method verified the above results. Conclusion This method is suitable for the rapid screening of common mutation sites in Chinese PKU patients