Rapid genotyping of common MeCP2 mutations with an electronic DNA microchip using serial differential hybridization.
Rapid genotyping of common MeCP2 mutations with an electronic DNA microchip using serial differential hybridization.
复制标题
使用串行差异杂交，通过电子 DNA 微芯片对常见 MeCP2 突变进行快速基因分型。
DOI：
10.1016/s1525-1578(10)60461-8
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Hoffman,EricP
中科院分区：
文献类型：
--
作者：
Thistlethwaite,WilliamA;Moses,LindaM;Hoffbuhr,KristenC;Devaney,JosephM;Hoffman,EricP
Rett syndrome is a neurodevelopmental disorder that affects females almost exclusively, and in which eight common point mutations on the X-linked MeCP2 gene are knows to cause over 70% of mutation-positive cases. We explored the use of a novel platform to detect the eight common mutations in Rett syndrome patients to expedite and simplify the process of identification of known genotypes. The Nanogen workstation consists of a two-color assay based on electric hybridization and thermal discrimination, all performed on an electronically active NanoChip. This genotyping platform was tested on 362 samples of a pre-determined genotype, which had been previously identified by a combination of DHPLC (denaturing high performance liquid chromatography) and direct sequencing. This genotyping technique proved to be rapid, facile, and displayed a specificity of 100% with 3% ambiguity. In addition, we present consecutive testing of seven mutations on a single pad of the NanoChip. This was accomplished by tagging down two amplimers together and serially hybridizing for seven different loci, allowing us to genotype samples for seven of the eight common Rett mutations on a single pad. This novel method displayed the same level of specificity and accuracy as the single amplimer reactions, and proved to be faster and more economical.
