PCR-ASO技术对苯丙氨酸羟化酶基因外显子突变的分析

The Analysis of Mutations of PAH Gene by PCR-ASO Technique

目的：检测苯丙酮尿症（ＰＫＵ）可疑患者的苯丙氨酸羟化酶（ＰＡＨ）基因外显子７、１１、１２（Ｅ７、１１、１２）区域的点突变。方法：应用ＰＣＲ方法对６例可疑ＰＫＵ患者的ＰＡＨ基因Ｅ７、１１、１２区域进行扩增，再用γ－３２Ｐ－ＡＴＰ标记的寡核苷酸探针（ＡＳＯ）杂交技术对其进行分析。结果：其中３例仅与含有Ｒ２４３Ｑ突变的异常探针杂交，不与正常探针杂交，说明其为Ｒ２４３Ｑ突变纯合子。结论：ＰＣＲ－ＡＳＯ探针可以特异地与ＰＡＨ突变位点杂交，是诊断ＰＫＵ的一种准确、可行的方法。

Objective:The purpose of this study was to detect the mutation in Exon 7 of phenylalanine hydroxylase (PAH) gene in 6 children suspected of phenyketonuria(PKU).Methods:The Exon 7 area of PAH gene in 6 suspected children was analyzed using PCR and γ-32-ATP labelled allele specific oligonucleotide(ASO) technique.Result:Homozygotes of Exon 7(R243Q) was found in 3 children. Conclusion:PCR-ASO technique can hybridize with PAH mutation specifically. It was an effective, available procedure in PKU genetic diagnosis.