一种用于检测脆性X智力低下1基因前突变和全突变等位基因的增强型聚合酶链反应检测法。
An enhanced polymerase chain reaction assay to detect pre- and full mutation alleles of the fragile X mental retardation 1 gene.
作者信息
Saluto Alessandro, Brussino Alessandro, Tassone Flora, Arduino Carlo, Cagnoli Claudia, Pappi Patrizia, Hagerman Paul, Migone Nicola, Brusco Alfredo
机构信息
Dipartimento di Genetica Biologia e Biochimica, Università degli Studi di Torino, via Santena 19, 10126 Torino, Italy.
出版信息
J Mol Diagn. 2005 Nov;7(5):605-12. doi: 10.1016/S1525-1578(10)60594-6.
Abstract
Several diagnostic strategies have been applied to the detection of FMR1 gene repeat expansions in fragile X syndrome. Here, we report a novel polymerase chain reaction-based strategy using the Expand Long Template PCR System (Roche Diagnostics, Mannheim, Germany) and the osmolyte betaine. Repeat expansions up to approximately 330 CGGs in males and up to at least approximately 160 CGGs in carrier women could be easily visualized on ethidium bromide agarose gels. We also demonstrated that fluorescence analysis of polymerase chain reaction products was a reliable tool to verify the presence of premutation and full mutation alleles both in males and in females. This technique, primarily designed to detect premutation alleles, can be used as a routine first screen for expanded FMR1 alleles.
摘要
几种诊断策略已应用于脆性X综合征中FMR1基因重复扩增的检测。在此，我们报告一种基于聚合酶链反应的新策略，该策略使用Expand Long Template PCR System（德国曼海姆罗氏诊断公司）和渗透剂甜菜碱。在溴化乙锭琼脂糖凝胶上可以轻松观察到男性中高达约330个CGG的重复扩增以及携带者女性中至少约160个CGG的重复扩增。我们还证明，聚合酶链反应产物的荧光分析是验证男性和女性中前突变和全突变等位基因存在的可靠工具。该技术主要设计用于检测前突变等位基因，可作为扩增FMR1等位基因的常规初步筛查方法。
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