title: Highly sensitive chemiluminescent point mutation detection by circular strand-displacement amplification reaction.
abstract: Single nucleotide polymorphism (SNP) genotyping is attracting extensive attentions owing to its direct connections with human diseases including cancers. Here, we have developed a highly sensitive chemiluminescence biosensor based on circular strand-displacement amplification and the separation by magnetic beads reducing the background signal for point mutation detection at room temperature. This method took advantage of both the T4 DNA ligase recognizing single-base mismatch with high selectivity and the strand-displacement reaction of polymerase to perform signal amplification. The detection limit of this method was 1.3 × 10(-16)M, which showed better sensitivity than that of most of those reported detection methods of SNP. Additionally, the magnetic beads as carrier of immobility was not only to reduce the background signal, but also may have potential apply in high through-put screening of SNP detection in human genome.
authors.affiliation: College of Biotechnology and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, PR China.
authors.forename: Chao
authors.initials: C
authors.lastname: Shi
authors.forename: Yujie
authors.initials: Y
authors.lastname: Ge
authors.forename: Hongxi
authors.initials: H
authors.lastname: Gu
authors.forename: Cuiping
authors.initials: C
authors.lastname: Ma
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D004247:DNA / Q000235:genetics*; D011088:DNA Ligases / Q000378:metabolism; D006801:Humans; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D017354:Point Mutation
chemical_list: D004247:DNA; D011088:DNA Ligases