A Novel Single Nucleotide Polymorphisms Detection Sensors Based on Magnetic Nanoparticles Array and Dual-Color Single Base Extension
A Novel Single Nucleotide Polymorphisms Detection Sensors Based on Magnetic Nanoparticles Array and Dual-Color Single Base Extension
复制标题
基于磁性纳米颗粒阵列和双色单碱基延伸的新型单核苷酸多态性检测传感器
DOI：
10.1166/jnn.2010.2386
复制
发表时间：
2010-08-01
影响因子：
--
通讯作者：
He, Nongyue
中科院分区：
文献类型：
--
作者：
Liu, Hongna;Li, Song;He, Nongyue
为满足大规模遗传学研究的需要，设计了一种基于金包被磁性纳米颗粒（GMNP）阵列和双色单碱基延伸的高通量、自动化SNPs基因分型方法。将生物素化的单碱基延伸引物捕获到用链霉亲和素包被的GMNP上。采用双色荧光单碱基延伸法作为SNPs鉴定的工具，然后将带有荧光团的GMNP点样到载玻片上，制成“微珠阵列”。利用该平台对12份样本的MTHFR基因C677 T位点进行了基因分型。结果表明，所有纯合子样品的信噪比均大于30，杂合子样品的荧光强度比值在1左右。该方法克服了背景扣除法的缺点，直接读取GMNP的荧光值来确定其基因型，不需要对FOR产物进行纯化和复杂的还原处理，具有操作简单、节省人力、灵敏度高和自动化程度高等优点。
To fulfill the increasing need for large-scale genetic research, a high-throughput and automated SNPs genotyping method based on gold coated magnetic nanoparticles (GMNPs) array and dual-color single base extension has been designed. Biotinylated single base extension primer were captured onto the GMNPs coated with streptavidin. Dual-color fluorescence single base extension was employed as a tool for SNPs identification, and then a "bead array" was fabricated by spotting GMNPs with fluorophore on the glass slide. Using this platform, MTHFR gene C677T locus from 12 samples was genotyped. The results showed that all homozygous samples gave a signal/noise ratio over 30, and the fluorescent intensities ratios of heterozygote samples were around 1. With advantages to defeat the background subtract feature and read-out the fluorescence values of the GMNPs directly to determine their genotypes, without the necessary procedures for purification and complex reduction of FOR products, the application of this strategy to large-scale SNP studies will be simple, labor-saving, high sensitive and potential for automation.