title: Sequence-Specific Biosensing of DNA Target through Relay PCR with Small-Molecule Fluorophore.
abstract: Polymerase chain reaction coupled with signal generation offers sensitive recognition of target DNA sequence; however, these procedures require fluorophore-labeled oligonucleotide probes and high-tech equipment to achieve high specificity. Therefore, intensive research has been conducted to develop reliable, convenient, and economical DNA detection methods. The relay PCR described here is the first sequence-specific detection method using a small-molecule fluorophore as a sensor and combines the classic 5'-3' exonuclease activity of Taq polymerase with an RNA mimic of GFP to build a label-free DNA detection platform. Primarily, Taq polymerase cleaves the 5' noncomplementary overhang of the target specific probe during extension of the leading primer to release a relay oligo to initiate tandem PCR of the reporting template, which encodes the sequence of RNA aptamer. Afterward, the PCR product is transcribed to mRNA, which could generate a fluorescent signal in the presence of corresponding fluorophore. In addition to high sensitivity and specificity, the flexibility of choosing different fluorescent reporting signals makes this method versatile in either single or multiple target detection.
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Afshan
authors.initials: A
authors.lastname: Yasmeen
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Feng
authors.initials: F
authors.lastname: Du
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Yongyun
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Juan
authors.initials: J
authors.lastname: Dong
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Haodong
authors.initials: H
authors.lastname: Chen
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Cui
authors.affiliation: Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences , Chengdu 610041, People's Republic of China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Tang
mesh_terms: D015374:Biosensing Techniques; D004247:DNA / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D005456:Fluorescent Dyes; D004247:DNA