The Functional Frame Nucleic Acid-mediated Rapid Detection Of IDH-1 Gene SNP In Glioma

Objective: The detection of single nucleotide polymorphism(SNP)of isocitrate dehydrogenase-1(IDH-1)is crucial for the diagnosis,staging and treatment of glioma.The purpose of this study is to design a rapid,sequencing-free detection method for IDH-1 SNP in glioma.Methods: In this study,the transcribed m RNA of IDH-1 gene was targeted for the detection of R132 H SNP.The detection assay based on the thermodynamic divergence of target binding process of the detection probe and the competitive probe.The functional frame nucleic acid(DNA tetrahedron here)was utilized to facilitate the binding of the probe to the detection surface,and to optimize the detection sensitivity.First,we designed the DNA tetrahedral nanoprobe and the detection assay.NUPACK was utilized to validate the detection assay in thermodynamic aspects.Second,DNA tetrahedral nanoprobe was synthesized.Homogenous reaction of the detection probes and the standard samples was then carried out to prove the feasibility of the detection assay.Last,the DNA tetrahedral nanoprobe was immobilized on the glass surface and the standard samples and the clinical samples were detected.Results: The feasibility of the proposed SNP detection assay was proved by the NUPACK algorithm and was further validated in the homogeneous reaction.The DNA tetrahedral nanoprobe was successfully synthesized and immobilized on the detection surface.In standard samples and clinical samples,significant differences were demonstrated between the wild type and mutation type of R132 H status of IDH-1 gene in surface detection.Conclusion: The detection assay proposed in this study can to a certain extent distinguish the mutation of the R132 H SNP of IDH-1 gene in standard samples and clinical samples effectively.