Zschocke J, Graham C A, Carson D J, Nevin N C
Department of Medical Genetics, Belfast City Hospital.
Am J Hum Genet. 1995 Dec;57(6):1311-7.
We present a multistep approach for the rapid analysis of phenylketonuria (PKU) mutations. In the first step, three common mutations and a polymorphic short tandem repeat (STR) system are rapidly analyzed with a fluorescent multiplex assay. In the second step, minihaplotypes combining STR and VNTR data are used to determine rare mutations likely to be present in an investigated patient, which are then confirmed by restriction enzyme analysis. The remaining mutations are analyzed with denaturant gradient-gel electrophoresis and sequencing. The first two steps together identify both mutations in 90%-95% of PKU patients, and results can be obtained within 2 d. We have investigated 121 Northern Irish families with hyperphenylalaninemia, including virtually all patients born since 1972, and have found 34 different mutations on 241 of the 242 mutant alleles. Three mutations (R408W, I65T, and F39L) account for 57.5% of mutations, while 14 mutations occur with a frequency of 1%-6%. The present analysis system is efficient and inexpensive and is particularly well suited to routine mutation analysis in a diagnostic setting.
我们提出了一种用于快速分析苯丙酮尿症（PKU）突变的多步骤方法。第一步，通过荧光多重分析法快速分析三种常见突变和一个多态性短串联重复序列（STR）系统。第二步，结合STR和VNTR数据的微型单倍型用于确定被调查患者可能存在的罕见突变，然后通过限制性酶切分析进行确认。其余突变则通过变性梯度凝胶电泳和测序进行分析。前两步共同鉴定出90%-95%的PKU患者的两种突变，且2天内即可获得结果。我们调查了121个患有高苯丙氨酸血症的北爱尔兰家庭，包括自1972年以来出生的几乎所有患者，在242个突变等位基因中的241个上发现了34种不同的突变。三种突变（R408W、I65T和F39L）占突变的57.5%，而14种突变的发生频率为1%-6%。本分析系统高效且廉价，特别适用于诊断环境中的常规突变分析。
