title: Sensitive SERS detection of DNA methyltransferase by target triggering primer generation-based multiple signal amplification strategy.
abstract: A novel and sensitive surface-enhanced Raman scattering (SERS) method is proposed for the assay of DNA methyltransferase (MTase) activity and evaluation of inhibitors by developing a target triggering primer generation-based multiple signal amplification strategy. By using of a duplex substrate for Dam MTase, two hairpin templates and a Raman probe, multiple signal amplification mode is achieved. Once recognized by Dam MTase, the duplex substrate can be cleaved by Dpn I endonuclease and two primers are released for triggering the multiple signal amplification reaction. Consequently, a wide dynamic range and remarkably high sensitivity are obtained under isothermal conditions. The detection limit is 2.57×10(-4)UmL(-1). This assay exhibits an excellent selectivity and is successfully applied in the screening of inhibitors for Dam MTase. In addition, this novel sensing system is potentially universal as the recognition element can be conveniently designed for other target analytes by changing the substrate of DNA MTase.
authors.affiliation: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
authors.forename: Chuanfeng
authors.initials: C
authors.lastname: Yu
authors.affiliation: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
authors.forename: Huixia
authors.initials: H
authors.lastname: Han
authors.affiliation: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China.
authors.forename: Caisheng
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, PR China. Electronic address: zhangxr7407@126.com.
authors.forename: Xiaoru
authors.initials: X
authors.lastname: Zhang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods; D057075:Enzyme Assays / Q000379:methods; D006801:Humans; D057230:Limit of Detection; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000032:analysis / Q000097:blood*; D013059:Spectrum Analysis, Raman / Q000379:methods*
chemical_list: C058693:Dam methyltransferase; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific)
keywords: DNA methyltransferase; Multiple signal amplification; Surface-enhanced Raman scattering; Target triggering primer generation