Samiotaki M, Kwiatkowski M, Parik J, Landegren U
Department of Medical Genetics, Beijer Laboratory, Uppsala University, Sweden.
Genomics. 1994 Mar 15;20(2):238-42. doi: 10.1006/geno.1994.1159.
Genetic screening for sequence variants associated with disease is assuming increasing importance in clinical medicine as well as in research. We describe an efficient method for such analyses, comprising a combination of practical features: (1) Amplified DNA samples are analyzed for their ability to serve as templates in standardized allele-specific ligation reactions between oligonucleotide probes; (2) Two allele-specific probes, differentially labeled with either of two lanthanide labels, compete for ligation to a third oligonucleotide (the signal from the two labeled probes can thus be directly compared in a sensitive time-resolved fluorescence detection reaction); and (3) Large sets of analyses are processed in parallel using a 96-pin capture manifold, serving to reduce pipetting steps and the risk of contamination. We present here the basis of the technique and its application to the screening for two common mutations causing cystic fibrosis and alpha 1-antiytrypsin deficiency.
对与疾病相关的序列变异进行基因筛查在临床医学和研究中都日益重要。我们描述了一种用于此类分析的高效方法，该方法具有以下实际特点：（1）分析扩增的DNA样本作为寡核苷酸探针之间标准化等位基因特异性连接反应模板的能力；（2）用两种镧系元素标签之一进行差异标记的两种等位基因特异性探针竞争与第三种寡核苷酸的连接（因此，在灵敏的时间分辨荧光检测反应中可以直接比较两种标记探针的信号）；（3）使用96针捕获阵列并行处理大量分析，以减少移液步骤和污染风险。我们在此介绍该技术的基础及其在筛查导致囊性纤维化和α1 -抗胰蛋白酶缺乏症的两种常见突变中的应用。
