Detection of Single Nucleotide Polymorphism (SNP) Variation of a Gene Sequence on Membrane-Based Lateral-Flow Strips

Abstract

This study used appropriate primers to distinguish the gene model, HLA-A31:01, on membrane-based lateral-flow (MBLF) strips from its allele, which is with an SNP. Using primers designed with a mismatch base on one or two sides next to the SNP spot was verified as a good approach. In the optimal condition, the detection limits of 1~0.1 ng/μL nucleotides were in agreement with reports in the literature, and the intra- and inter-assay tests ensured the detection reproducibility of this approach with CV% of 2.5~15.9% and 1.7~14.7%, respectively. The detection specificity was also validated by the tests on the selected negative-control genes. The tests on MBLF strips in this study showed an easy, robust, reproducible, and reliable detection methodology for untrained personnel at care points with limited instrument and particularly for avoiding medications from faulty prescriptions.

Keywords

• single nucleotide polymorphism (SNP)
• membrane-based lateral-flow (MBLF) strips
• polymerase chain reaction (PCR)
• primer design
• biosensors