Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchips

被引:184
作者
Gilles, PN
Wu, DJ
Foster, CB
Dillon, PJ
Chanock, SJ
机构
[1] Nanogen Inc, San Diego, CA 92121 USA
[2] NCI, Pediat Oncol Branch, NIH, Bethesda, MD 20892 USA
关键词
single nucleotide polymorphism; mannose binding protein; microarray; chip;
D O I
10.1038/7921
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have developed a rapid assay for single nucleotide polymorphism (SNP) detection that utilizes electronic circuitry on silicon microchips. The method was validated by the accurate discrimination of blinded DNA samples for the complex quadra-allelic SNP of mannose binding protein. The microchip directed the transport, concentration, and attachment of amplified patient DNA to selected electrodes (test sites) creating an array of DNA samples. Through control of the electric field, the microchip enabled accurate genetic identification of these samples using fluorescently labeled DNA reporter probes. The accuracy of this approach was established by internal controls of dual labeled reporters and by using mismatched sequences in addition to the wild-type and variant reporter sequences to validate the SNP-genotype. The ability to customize this assay for multiple genes has advantages over other existing approaches.
引用
收藏
页码:365 / 370
页数:6
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