title: A label-free aptasensor for highly sensitive detection of ATP and thrombin based on metal-enhanced PicoGreen fluorescence.
abstract: A label-free fluorescence aptasensor for highly selective and sensitive detection of ATP and thrombin was developed by using PicoGreen (PG) as signal molecule and surface-bound metal-enhanced fluorescence (MEF) substrates (silver island films, SIFs) as signal enhancers. On binding with ATP or thrombin, aptamers undergo structure switching, leading to a reduction of fluorescence intensity of PG. Chang of fluorescence intensity can be magnified by SIFs. The limit of detection for ATP and thrombin is 1.3 nM and 0.073 nM, respectively. The fluorescence quenching efficiency is linear in the logarithmic scale with ATP concentration range from 10 nM to 100 μM (R(2)=0.995) and thrombin concentration range from 0.1 nM to 100 nM (R(2)=0.997). The coefficients of variation of the intra-assay reproducibility and inter-assay reproducibility for ATP (10 μM) assay are 3.8% and 5.2%, respectively. In addition, the aptasensor is stable and can be reliably used for ATP measurement in biological samples. Overall, the aptasensor can be a useful and cost effective tool for the specific detection of ATP, thrombin and potentially other biomolecules in biological samples.
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China; Department of Laboratory Medicine, Fuzong Clinical College, Fujian Medical University, Fuzhou 350025, PR China.
authors.forename: Kaiyu
authors.initials: K
authors.lastname: Wang
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Liao
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China.
authors.forename: Xiangyue
authors.initials: X
authors.lastname: Yang
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Laboratory Medicine, Fuzong Clinical College, Fujian Medical University, Fuzhou 350025, PR China.
authors.forename: Weirong
authors.initials: W
authors.lastname: Yao
authors.affiliation: Center for Research at the Bio/Nano Interface, Department of Chemistry and Shands Cancer Center, University of Florida, Gainesville, Florida 32611, United States; Plant Pathology Lab, North Florida Research and Education Center, University of Florida, Quincy, Florida 32351, United States; Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha 410082, PR China. Electronic address: tan@chem.ufl.edu.
authors.forename: Weihong
authors.initials: W
authors.lastname: Tan
authors.affiliation: Institute of Laboratory Medicine, Fuzhou General Hospital of Nanjing Military Command, Fuzhou 350025, PR China; Department of Laboratory Medicine, Dongfang Hospital, Xiamen University, Fuzhou 350025, PR China; Department of Laboratory Medicine, Fuzong Clinical College, Fujian Medical University, Fuzhou 350025, PR China. Electronic address: lanxp@sina.com.
authors.forename: Xiaopeng
authors.initials: X
authors.lastname: Lan
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation*; D045424:Complex Mixtures / Q000032:analysis; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry; D009930:Organic Chemicals / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012834:Silver / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000295:instrumentation* / Q000379:methods; D013194:Staining and Labeling; D013917:Thrombin / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D045424:Complex Mixtures; D005456:Fluorescent Dyes; D009930:Organic Chemicals; C099476:PicoGreen; D012834:Silver; D000255:Adenosine Triphosphate; D013917:Thrombin
keywords: ATP; Aptasensor; Label-free; Metal-enhanced fluorescence; PicoGreen; Thrombin