title: A novel lateral flow assay based on GoldMag nanoparticles and its clinical applications for genotyping of MTHFR C677T polymorphisms.
abstract: 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.
authors.affiliation: College of Life Sciences, Northwest University, Xi'an, 710069, China. yalicui@nwu.edu.cn.
authors.forename: Wenli
authors.initials: W
authors.lastname: Hui
authors.forename: Sinong
authors.initials: S
authors.lastname: Zhang
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.forename: Yinsheng
authors.initials: Y
authors.lastname: Wan
authors.forename: Juanli
authors.initials: J
authors.lastname: Zhu
authors.forename: Gang
authors.initials: G
authors.lastname: Zhao
authors.forename: Songdi
authors.initials: S
authors.lastname: Wu
authors.forename: Dujuan
authors.initials: D
authors.lastname: Xi
authors.forename: Qinlu
authors.initials: Q
authors.lastname: Zhang
authors.forename: Ningning
authors.initials: N
authors.lastname: Li
authors.forename: Yali
authors.initials: Y
authors.lastname: Cui
mesh_terms: D005260:Female; D060005:Genotyping Techniques / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D008297:Male; D053768:Metal Nanoparticles / Q000737:chemistry*; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide
chemical_list: D006046:Gold; C573423:MTHFR protein, human; D042965:Methylenetetrahydrofolate Reductase (NADPH2)