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.

A PCR-lateral flow assay system based on gold magnetic nanoparticles for CYP2C19 genotyping and its clinical applications

“…It shows great potential for serving as diagnostic or therapeutic agents in a variety of medical fields [18][19][20]. In a previous study, our laboratory established a novel lateral flow assay (LFA) system assembled with GMNPs for the visual detection of MTHFR C677T polymorphism which relies on the hybridization reaction on test line [21]. This method is simple, accurate, affordable, and user-friendly for SNP detection.…”

Rapid Detection of Anti-SARS-CoV-2 IgM and IgG Using a Selenium Nanoparticle-based Lateral Flow Immunoassay

“…LFIAs utilize various types of labeling probes that either provide quantitative detection or qualitative detection [11]. Quantitative detection labeling probes include uorescent microspheres [12], upconversion luminescence [13], quantum dots [14,15], and magnetic particles [16]; qualitative detection labeling probes include colloidal gold [17], selenium nanoparticle [18], and nanocarbon [11].…”

Sensitive genotyping of mutations in the EGFR gene from NSCLC patients using PCR-GoldMag lateral flow device

“…Amplification refractory mutation system (ARMS)-PCR is a simple and accurate method that could discriminate between mutant and wild-type DNA. GoldMag lateral flow device combined with ARMS-PCR was set up as a simple and rapid genotyping method for methylenetetrahydrofolate reductase ( MTHFR ) C677T 21 . The new PCR-GoldMag lateral flow device for SNP detection, including MTHFR C677T and Apolipoprotein E polymorphisms, has been validated by sequencing for more than 2,000 genomic DNAs in 6 hospitals in China, and showed a high specificity and sensitivity 21 , 22 .…”

“…GoldMag lateral flow device combined with ARMS-PCR was set up as a simple and rapid genotyping method for methylenetetrahydrofolate reductase ( MTHFR ) C677T 21 . The new PCR-GoldMag lateral flow device for SNP detection, including MTHFR C677T and Apolipoprotein E polymorphisms, has been validated by sequencing for more than 2,000 genomic DNAs in 6 hospitals in China, and showed a high specificity and sensitivity 21 , 22 . This method allows for rare signals to be detected with greater sensitivity, tends to be faster and cheaper, and hence can be used as “targeted method” for genotyping of EGFR gene.…”