Chehab F F, Wall J
Department of Laboratory Medicine, University of California, San Francisco 94143-0134.
Hum Genet. 1992 May;89(2):163-8. doi: 10.1007/BF00217117.
We describe the implementation of a modified version of the reverse dot blot hybridization technology to detect eight cystic fibrosis mutations. The method is simple, quick, reliable, inexpensive, and nonradioactive and utilizes the sensitivity of the polymerase chain reaction coupled with colored or chemiluminescent substrates for mutation detection. We have used this system in a clinical laboratory to identify the delta F508, G542X, G551D, R553X, 621 + 1G----T, W1282X, N1303K, and 1717G----A mutations. The technique is practical for genotyping individuals at many potential mutation sites, as in cystic fibrosis and beta-thalassemia, in which over 95 mutations can cause disease. This technology appears to be the method of choice for the widespread carrier screening of multiple cystic fibrosis mutations.
我们描述了一种改良的反向斑点杂交技术的应用，用于检测八种囊性纤维化突变。该方法简单、快速、可靠、成本低廉且无放射性，利用聚合酶链反应的敏感性，并结合显色或化学发光底物进行突变检测。我们已在临床实验室中使用该系统来鉴定ΔF508、G542X、G551D、R553X、621 + 1G→T、W1282X、N1303K和1717G→A突变。该技术对于在许多潜在突变位点对个体进行基因分型很实用，如在囊性纤维化和β地中海贫血中，超过95种突变可导致疾病。这项技术似乎是广泛筛查多种囊性纤维化突变携带者的首选方法。
