Single Nucleotide Polymorphism Of Nucleic Acid Detection Technology In The Test Strip

Objective: Developing a rapid method for detection of single nucleotide polymorphisms and mutation.Methods: We first reported proof-of-principle for a genotyping assay approach that can detect single nucleotide polymorphisms (SNPs) and single base mutation by the combination of Nucleic acid Lateral-Flow Diagnostic Strip, PCR and the ligase detection reaction. This assay provided a convenient and powerful detection that enabled to detect single-base discrimination. The assay could be implemented via three steps:Amplification: PCR amplified the target sequence;Ligation: After two detection probes hybridized to the target DNA, the ligase reaction was triggered when the perfect match had happened;Detection: When the reaction mixture was heated with 0.05M NaOH to denature the double strand DNA, only the ligated oligonucleotides would be detected by the strip.Results:The approach has been demonstrated by 50 identifications of a single-base mutation in HBV YMDD region which can cause the resistance to the Lamivudine. It showed same genotypes as results of DNA sequencing and others.Conclusion: The assay is accurate, simple, rapid and inexpensive. It could have great potentials in applications related SNP/mutation detection in clinics and hospitals.