Picci L, Anglani F, Scarpa M, Zacchello F
Department of Pediatrics, University of Padua, Italy.
Hum Genet. 1992 Mar;88(5):552-6. doi: 10.1007/BF00219343.
We have developed a simple rapid DNA screening test that allows us simultaneously to analyze seven CF mutations (delta F508, R347P, S549N, G551D, R553X, R334W, 444delA) that together account for about 60% of all CF mutations in the Italian population. It consists of three steps: multiplex polymerase chain reaction (PCR) amplification of exons 4, 7, 10 and 11; restriction endonuclease digestion of the PCR products; and vertical polyacrylamide gel electrophoresis analysis. We have used our multiplex assay for analyzing 15 CF chromosomes (non delta F508) and have found 3 cases of the R553X mutation; the latter have been confirmed by amplification and digestion of exon 11.
我们开发了一种简单快速的DNA筛查检测方法，该方法能让我们同时分析7种囊性纤维化（CF）突变（ΔF508、R347P、S549N、G551D、R553X、R334W、444delA），这些突变加起来约占意大利人群中所有CF突变的60%。它包括三个步骤：外显子4、7、10和11的多重聚合酶链反应（PCR）扩增；PCR产物的限制性内切酶消化；以及垂直聚丙烯酰胺凝胶电泳分析。我们已使用我们的多重检测方法分析了15条CF染色体（非ΔF508），并发现了3例R553X突变病例；后者已通过外显子11的扩增和消化得到证实。
