Research And Application Of Drug Related Gene Mutation Detectuon Method

Single nucleotide polymorphisms(SNP)has been considered to be the important reason for the complexity and diversity in gene.?1-adrenergic receptor gene(ADRB1)SNP can directly influence the antihypertensive effect of metoprolol with hypertension.A polymorphism of Vitamin K epoxide reductase complex subunit 1(VKORC1)is associated with warfarin dosing in people.Thus,the establishment of a new SNP molecular detection method can better understand the individual differences in response to a particular drug.Therefore,achieved individualized treatment goal,and reduce medical and medical costs.In this paper,different nano-materials modified electrode was used to achieve the detection of two common clinical applied drug-related SNP,to accurate guidance of individualized clinical medication.The research is divided into the following two parts: 1.Ultrasensitive detection of the ?1-adrenergic receptor gene mutation based on Graphene oxide/ Cerium oxide/ platinum nanoparticles as a lableIn this study,a specific assay based on graphene oxide/cerium oxide/ platinum nanoparticles(GO/CeO2/Pt NPs)as a beacon,gold nanoparticles/ streptavidin(AuNPs/SA)as the electrode modified nanomaterial for the detection of ADRB1 mutation in serum was constructed.CeO2 was first immobilized on the GO,and then PtNPs was in situ reductions on the surface of CeO2.As a result,GO/CeO2/PtNPs were synthesized and obtained the favorable catalytic performance.After the depositing of AuNPs on the surface of the glassy carbon electrode(GCE),the SA was modified to link the biotin-labeled probe.Under the optimal experimental conditions,the prepared biosensor had high sensitivity,good stability and wide liner range from 1fM to 100 fM and 100 fM to 10 nM,with a low detection limit of 0.33 fM.In addition,the recovery rate is from 99.4% to 101.9%,indicating that the biosensor can be used in serum ADRB1 gene mutation detection.2.An electrochemicial biosensor for the ultrasensitive detection of VKORC1 genotyping with C60NPs-based nanoparticles-assisted signal amplificationIn this work,an electrochemical biosensor was fabricated by fullerene-amino terminated polyamidoamine dendrimer-platinum porous nanoparticles(C60NPs-PAMAM-Pt PNPs)nanocomposites serving as a signal probe and reduced graphene oxide–tetraethylene pentamine/gold nanoparticles/streptavidin(rGO-TEPA/AuNPs/SA)serving as the electrode modified nanomaterial for the selective detection of VKORC1 mutant gene.In this design,C60 NPs not only have redox activity,but also act as the biomolecule carrier to immobilize PtPNPs to obtain C60NPs-PAMAM-PtPNPs.rGO-TEPA was dropped onto the GCE surface,then the prepared electrode was modified with AuNPs by electrodeposition and SA was added to link the biotin-labeled tetrahedral DNA.Under the optimal conditions,the prepared biosensor exhibited high sensitivity,good selectivity and reproducibility,a wide linear range from 1 pM to 10 nM and a low detection limit of 33 fM.In addition,the recovery rate of the prepared sensor was 96.7-103.94%,indicating its potential use in serum VKORC1 gene mutation detection.