Liu Hongna, Li Song, Ji Meiju, Nie Libo, Chen Jianrong, Miao Yuqing, He Nongyue
Hunan Key Laboratory of Green Packaging and Application of Biological Nanotechnology, Hunan University of Technology, Zhuzhou 412008, China.
J Nanosci Nanotechnol. 2008 Jan;8(1):405-9.
We have developed a novel approach to fabricate single nucleotide polymorphisms (SNPs) library on magnetic nanoparticles (MNPs) based on adaptor PCR. Each SNP locus in the library was interrogated by hybridization with a pair of allele specific dual-color fluorescence (Cy3, Cy5) probes to determine SNP. Two SNPs loci (M235T and A-6G) associated with essential hypertension in the angiotensinogen (AGT) gene were detected by this method and their fluorescent signals were quantified. The fluorescent ratios (match probe: mismatch probe signal) of homozygous genotypes were over 3.0, whereas heterozygous genotypes had ratios near to 1.0. Without any complex multiplex PCR procedure, it is a simple, efficient and reliable method for the multiplex SNPs detection using limited amount of DNA samples from individuals.
我们开发了一种基于衔接子聚合酶链反应（PCR）在磁性纳米颗粒（MNPs）上构建单核苷酸多态性（SNP）文库的新方法。通过与一对等位基因特异性双色荧光（Cy3、Cy5）探针杂交来检测文库中的每个SNP位点，以确定SNP。用该方法检测了血管紧张素原（AGT）基因中与原发性高血压相关的两个SNP位点（M235T和A-6G），并对其荧光信号进行了定量。纯合基因型的荧光比率（匹配探针：错配探针信号）超过3.0，而异合子基因型的比率接近1.0。该方法无需复杂的多重PCR程序，是一种使用个体有限DNA样本进行多重SNP检测的简单、高效且可靠的方法。
