A novel lateral flow assay based on GoldMag nanoparticles and its clinical applications for genotyping of MTHFR C677T polymorphisms
A novel lateral flow assay based on GoldMag nanoparticles and its clinical applications for genotyping of MTHFR C677T polymorphisms
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DOI：
10.1039/c5nr07547e
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发表时间：
2016-02-14
期刊：
影响因子：
6.7
通讯作者：
Cui, Yali
中科院分区：
文献类型：
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作者：
Hui, Wenli;Zhang, Sinong;Cui, Yali
目前的单核苷酸多态性（SNP）检测技术需要繁琐的实验程序和昂贵而复杂的仪器。本研究将ARMS-PCR与基于金磁纳米颗粒（GoldMag）的侧向流分析（LFA）相结合，成功建立了一种可视化基因分型方法，并应用于亚甲基四氢叶酸还原酶（MTHFR）C677 T的基因分型。MTHFR基因的C677 T取代导致疾病风险增加。将PCR产物加载到LFA装置上后5分钟内，通过目视观察可轻松获得基因分型结果。该系统能够准确评估每个测试样品从5 ng到1200 ng的初始起始基因组DNA量的宽检测范围。方法的最低检测限为5 ng。此外，我们的PCR-LFA系统被应用于临床试验，筛选1721个人的C677 T基因型。PCR-LFA基因分型结果与测序结果的符合率达99.6%。总的来说，我们的PCR-LFA已被证明是快速，准确，灵敏和廉价的。该方法适用于C677 T SNP的实验室和临床筛查。更有希望的是，它也可以扩展到检测其他基因的SNP。
Current techniques for single nucleotide polymorphism (SNP) detection require tedious experimental procedures and expensive and sophisticated instruments. In this study, a visual genotyping method has been successfully established via combining ARMS-PCR with gold magnetic nanoparticle (GoldMag)based lateral flow assay (LFA) and applied to the genotyping of methylenetetrahydrofolate reductase (MTHFR) C677T. C677T substitution of the gene MTHFR leads to an increased risk of diseases. The genotyping result is easily achievable by visual observation within 5 minutes after loading of the PCR products onto the LFA device. The system is able to accurately assess a broad detection range of initial starting genomic DNA amounts from 5 ng to 1200 ng per test sample. The limit of detection reaches 5 ng. Furthermore, our PCR-LFA system was applied to clinical trials for screening 1721 individuals for the C677T genotypes. The concordance rate of the genotyping results detected by PCR-LFA was up to 99.6% when compared with the sequencing results. Collectively, our PCR-LFA has been proven to be rapid, accurate, sensitive, and inexpensive. This new method is highly applicable for C677T SNP screening in laboratories and clinical practices. More promisingly, it could also be extended to the detection of SNPs of other genes.