He Nongyue, Li Song, Liu Hongna
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China.
Methods Mol Biol. 2009;578:393-402. doi: 10.1007/978-1-60327-411-1_24.
A microarray-based method for detecting single nucleotide polymorphisms (SNPs) using solid-phase polymerase chain reaction (PCR) on magnetic nanoparticles (MNPs) was developed. In this method one primer with a biotin label is captured by streptavidin-coated MNPs (SA-MNPs), and the PCR products are directly amplified on the surface of SA-MNPs in a 96-well plate. The samples are further probed by hybridization with a pair of dual-color probes to determine SNP. The genotype of each sample can be simultaneously identified by scanning the microarray printed with the denatured fluorescent probes. All the reactions can be performed in the same reaction volume without the necessity of purification of intermediates. This approach represents a novel, simple, and labor-saving method for SNP genotyping and can be applied in automation system(s) to achieve high-throughput SNP detection.
开发了一种基于微阵列的方法，用于在磁性纳米颗粒（MNP）上使用固相聚合酶链反应（PCR）检测单核苷酸多态性（SNP）。在该方法中，带有生物素标签的一种引物被链霉亲和素包被的磁性纳米颗粒（SA-MNP）捕获，PCR产物在96孔板中的SA-MNP表面直接扩增。通过与一对双色探针杂交进一步检测样品以确定SNP。通过扫描印有变性荧光探针的微阵列可同时鉴定每个样品的基因型。所有反应都可以在相同反应体积中进行，无需纯化中间体。这种方法代表了一种用于SNP基因分型的新颖、简单且省力的方法，可应用于自动化系统以实现高通量SNP检测。
