Fluorescent aptasensor for antibiotic detection using magnetic bead composites coated with gold nanoparticles and a nicking enzyme.

Antibiotic abuse has been bringing serious pollution in water, which is closely related to human health. It is desirable to develop a new strategy for antibiotic detection. To address this problem, a sensitive fluorescent aptasensor for antibiotic detection was developed by utilizing gold nanoparticles modified magnetic bead composites (AuNPs/MBs) and nicking enzyme. AuNPs/MBs were synthesized with the help of polyethylenimine (PEI). The prepared AuNPs/MBs acted as dual-functional scaffolds that owned excellent magnetic separation capacity and strong covalent bio-conjugation. The non-specifically absorbed aptamers in AuNPs/MBs were less than that in MBs. Hence, the fluorescent aptasensor based on AuNPs/MBs show a better signal to background ratio than that based on carboxyl modified magnetic beads (MBs). In this work, ampicillin was employed as a model analyte. In the presence of ampicillin, the specific binding between ampicillin and aptamer induced structure-switching that led to the release of partial complementary DNA (cDNA) of aptamer. Then, the released cDNA initiated the cycle of nicking enzyme assisted signal amplification (NEASA). Therefore, a large amount of taqman probes were cleaved and fluorescence signal was amplified. The prepared fluorescent aptasensor bring sensitive detection in range of 0.1-100 ng mL-1 with the limit of detection of 0.07 ng mL-1. Furthermore, this aptasensor was also successfully applied in real sample detection with acceptable accuracy. The fluorescent aptasensor provides a promising method for efficient, rapid and sensitive antibiotic detection.

[1]  Jinsub Choi,et al.  Preparation and characterization of chemically functionalized silica-coated magnetic nanoparticles as a DNA separator. , 2009, The journal of physical chemistry. B.

[2]  Wei Xu,et al.  Ultrasensitive and selective colorimetric DNA detection by nicking endonuclease assisted nanoparticle amplification. , 2009, Angewandte Chemie.

[3]  Xiliang Luo,et al.  Signal amplified strategy based on target-induced strand release coupling cleavage of nicking endonuclease for the ultrasensitive detection of ochratoxin A. , 2013, Biosensors & bioelectronics.

[4]  H. Santos,et al.  Novel nanocomposites based on a strawberry-like gold- coated magnetite (Fe@Au) for protein separation in multiple myeloma serum samples , 2015, Nano Research.

[5]  Ultrasensitive voltammetric determination of kanamycin using a target-triggered cascade enzymatic recycling couple along with DNAzyme amplification , 2017, Microchimica Acta.

[6]  C. Fan,et al.  Isothermal Amplification of Nucleic Acids. , 2015, Chemical reviews.

[7]  P. He,et al.  A sensitive DNA electrochemical biosensor based on magnetite with a glassy carbon electrode modified by muti-walled carbon nanotubes in polypyrrole , 2005 .

[8]  Y. Duan,et al.  Plasma enhanced label-free immunoassay for alpha-fetoprotein based on a U-bend fiber-optic LSPR biosensor , 2015 .

[9]  B. B. Finlay,et al.  Effects of antibiotics on human microbiota and subsequent disease. , 2014, Annual review of microbiology.

[10]  Yu Wang,et al.  Target-aptamer binding triggered quadratic recycling amplification for highly specific and ultrasensitive detection of antibiotics at the attomole level. , 2015, Chemical communications.

[11]  Yue Gu,et al.  DNA Nanostructure-Based Magnetic Beads for Potentiometric Aptasensing. , 2015, Analytical chemistry.

[12]  Lianying Zhang,et al.  Efficient purification of lysozyme from egg white by 2-mercapto-5-benzimidazolesulfonic acid modified Fe3O4/Au nanoparticles. , 2016, Materials science & engineering. C, Materials for biological applications.

[13]  Longhua Tang,et al.  Enzyme-guided plasmonic biosensor based on dual-functional nanohybrid for sensitive detection of thrombin. , 2015, Biosensors & bioelectronics.

[14]  Da Xing,et al.  Sensitive and homogeneous protein detection based on target-triggered aptamer hairpin switch and nicking enzyme assisted fluorescence signal amplification. , 2012, Analytical chemistry.

[15]  Liuting Mo,et al.  Aptamer-integrated DNA nanostructures for biosensing, bioimaging and cancer therapy. , 2016, Chemical Society reviews.

[16]  Joseph Irudayaraj,et al.  Magnetic and gold-coated magnetic nanoparticles as a DNA sensor. , 2006, Analytical chemistry.

[17]  Changill Ban,et al.  Aptasensor for ampicillin using gold nanoparticle based dual fluorescence–colorimetric methods , 2012, Analytical and Bioanalytical Chemistry.

[18]  L. Toppare,et al.  A Novel Acetylcholinesterase Biosensor: Core-Shell Magnetic Nanoparticles Incorporating a Conjugated Polymer for the Detection of Organophosphorus Pesticides. , 2016, ACS applied materials & interfaces.

[19]  Anthony D. Keefe,et al.  Aptamers as therapeutics , 2010, Nature Reviews Drug Discovery.

[20]  L. Durso,et al.  Impacts of antibiotic use in agriculture: what are the benefits and risks? , 2014, Current opinion in microbiology.

[21]  Boris Mizaikoff,et al.  Piezoelectric sensors using molecularly imprinted nanospheres for the detection of antibiotics , 2016 .

[22]  Chunhong Zhu,et al.  Magnetic sensing film based on Fe₃O₄@Au-GSH molecularly imprinted polymers for the electrochemical detection of estradiol. , 2016, Biosensors & bioelectronics.

[23]  Yixiang Duan,et al.  An aptamer based method for small molecules detection through monitoring salt-induced AuNPs aggregation and surface plasmon resonance (SPR) detection , 2016 .

[24]  Mark R. Servos,et al.  Instantaneous and quantitative functionalization of gold nanoparticles with thiolated DNA using a pH-assisted and surfactant-free route. , 2012, Journal of the American Chemical Society.

[25]  Y. Duan,et al.  Chemistry, biology, and medicine of fluorescent nanomaterials and related systems: new insights into biosensing, bioimaging, genomics, diagnostics, and therapy. , 2014, Chemical reviews.

[26]  Feng Li,et al.  Highly sensitive homogeneous electrochemical aptasensor for antibiotic residues detection based on dual recycling amplification strategy. , 2016, Biosensors & bioelectronics.

[27]  C. Ban,et al.  Gold nanoparticle-based colorimetric detection of kanamycin using a DNA aptamer. , 2011, Analytical biochemistry.

[28]  Clara Pereira,et al.  Highly Monodisperse Fe3O4@Au Superparamagnetic Nanoparticles as Reproducible Platform for Genosensing Genetically Modified Organisms , 2016 .

[29]  Ning Gan,et al.  An ultrasensitive fluorescence aptasensor for chloramphenicol based on FRET between quantum dots as donor and the magnetic SiO2@Au NPs probe as acceptor with exonuclease-assisted target recycling , 2016 .

[30]  Rong Huang,et al.  Dual-mode protein detection based on Fe3O4-Au hybrid nanoparticles , 2012, Nano Research.

[31]  E. Hornes,et al.  Assessment of methods for covalent binding of nucleic acids to magnetic beads, Dynabeads, and the characteristics of the bound nucleic acids in hybridization reactions. , 1988, Nucleic acids research.

[32]  Y. Duan,et al.  Novel signal-enhancing immunoassay for ultrasensitive biomarker detection based on laser-induced fluorescence. , 2015, Analytical chemistry.

[33]  Lijie Cao,et al.  Cocaine detection via rolling circle amplification of short DNA strand separated by magnetic beads. , 2011, Biosensors & bioelectronics.

[34]  Huixiang Li,et al.  Label-free colorimetric detection of specific sequences in genomic DNA amplified by the polymerase chain reaction. , 2004, Journal of the American Chemical Society.

[35]  Min-Gon Kim,et al.  Novel antibody/gold nanoparticle/magnetic nanoparticle nanocomposites for immunomagnetic separation and rapid colorimetric detection of Staphylococcus aureus in milk. , 2013, Biosensors & bioelectronics.

[36]  S. Manzetti,et al.  The environmental release and fate of antibiotics. , 2014, Marine pollution bulletin.

[37]  Y. Duan,et al.  Amplified fluorescent aptasensor through catalytic recycling for highly sensitive detection of ochratoxin A. , 2015, Biosensors & bioelectronics.

[38]  S. M. Taghdisi,et al.  A selective and sensitive fluorescent aptasensor for detection of kanamycin based on catalytic recycling activity of exonuclease III and gold nanoparticles , 2016 .

[39]  Rui Li,et al.  Colorimetric determination of copper(II) using a polyamine-functionalized gold nanoparticle probe , 2015, Microchimica Acta.

[40]  Amplified binding-induced homogeneous assay through catalytic cycling of analyte for ultrasensitive protein detection. , 2016, Chemical communications.

[41]  M. Sadjadi,et al.  Fabrication and spectroscopic studies of folic acid-conjugated Fe3O4@Au core-shell for targeted drug delivery application. , 2015, Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy.

[42]  Qixing Zhou,et al.  Trends in antibiotic resistance genes occurrence in the Haihe River, China. , 2010, Environmental science & technology.

[43]  H. Ju,et al.  Chemiluminescence imaging for microRNA detection based on cascade exponential isothermal amplification machinery. , 2016, Analytica chimica acta.

[44]  Po-Jung Jimmy Huang,et al.  Dissociation and degradation of thiol-modified DNA on gold nanoparticles in aqueous and organic solvents. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[45]  Ailiang Chen,et al.  Replacing antibodies with aptamers in lateral flow immunoassay. , 2015, Biosensors & bioelectronics.

[46]  Wei Wei,et al.  Flow injection chemiluminescence immunoassay of microcystin-LR by using PEI-modified magnetic beads as capturer and HRP-functionalized silica nanoparticles as signal amplifier. , 2013, The Analyst.

[47]  Yuzhong Zhang,et al.  Ultrasensitive Multiplexed Immunoassay for Tumor Biomarkers Based on DNA Hybridization Chain Reaction Amplifying Signal. , 2016, ACS applied materials & interfaces.

[48]  Ming Zhou,et al.  Bioelectrochemical interface engineering: toward the fabrication of electrochemical biosensors, biofuel cells, and self-powered logic biosensors. , 2011, Accounts of chemical research.

[49]  A. Phulukdaree,et al.  Biosynthesis and computational analysis of amine-ended dual thiol ligand functionalized gold nanoparticles for conventional spectroscopy detection of melamine. , 2017, Journal of photochemistry and photobiology. B, Biology.

[50]  Yu Wang,et al.  A novel sandwich-type electrochemical aptasensor based on GR-3D Au and aptamer-AuNPs-HRP for sensitive detection of oxytetracycline. , 2017, Biosensors & bioelectronics.

[51]  W. Tan,et al.  Aptamer-conjugated optical nanomaterials for bioanalysis , 2012 .

[52]  Hui Li,et al.  An ultrasensitive homogeneous aptasensor for kanamycin based on upconversion fluorescence resonance energy transfer. , 2014, Biosensors & bioelectronics.

[53]  I. Willner,et al.  Application of the Hybridization Chain Reaction on Electrodes for the Amplified and Parallel Electrochemical Analysis of DNA , 2016 .

[54]  Oliver Reiser,et al.  Polymer- and dendrimer-coated magnetic nanoparticles as versatile supports for catalysts, scavengers, and reagents. , 2014, Accounts of chemical research.

[55]  Yuan Zhang,et al.  Determination of formaldehyde in blood plasma by high-performance liquid chromatography with fluorescence detection. , 2001, Journal of chromatography. B, Biomedical sciences and applications.

[56]  Yang Song,et al.  Nicking enzyme-assisted biosensor for Salmonella enteritidis detection based on fluorescence resonance energy transfer. , 2014, Biosensors & bioelectronics.