MicroRNAs (miRNAs) are considered as being promising biomarkers for hematological malignancies, their aging, progression and prognosis. The authors have developed a method for the detection of miRNA-155 by using surface plasmon resonance (SPR) imaging coupled to a nucleic acid-based amplification strategy using gold nanoparticles (AuNPs). The target miRNA-155 is captured by surface-bound DNA probes. After hybridization, DNA-AuNP are employed for signal amplification via DNA sandwich assembly, resulting in a large increase in the SPR signal. This method can detect miRNA-155 in concentrations down to 45 pM and over dynamic that extends from 50 pM to 5 nM. The assay is highly specific and can discriminate even a single base mismatch. It also is reproducible, precise, and was successfully applied to the determination of miRNA-155 in spiked real samples where it gave recoveries in the range between 86% and 98%. This biosensor provides an alternative approach for miRNA detection in biomedical research and clinical diagnosis, which is highly effective and efficient.
Graphical abstract Schematic of a surface plasmon resonance imaging biosensor for detection of miRNA-155 using strand displacement amplification and gold nanoparticle.
The authors describe an electrochemical method for the determination of the blood clotting factor IX (FIX). A nanogapped dielectrode (with a < 100 nm junction) was modified with antibody against FIX, and the resulting system was characterized by both impedance spectroscopy and voltammetry. In order to attain the improved sensitivity, gold nanoparticles were electrostatically attached to FIX. The current to voltage (I-V) measurement was carried out from 1 V to 5 V, where the entire calibration plot with 5 V was taken. This results in a limit of detection as low of 1 pM, which is much lower compared to the real concentration of FIX (87 nM) in human blood serum. The analytical range extends from 1 pM to 0.1 μM. The electrode is highly specific over other serum proteins.
Graphical abstract A nanogapped dielectrode with a <100 nm junction was modified with antibody against blood clotting factor IX and characterized by voltammetry. Gold nanoparticles were electrostatically attached to the analyte (blood clotting factor IX), and current/voltage (I-V) measurements were performed. Factor IX can be quantified with a limit of detection as low as 10 pM.
The authors describe a novel assay for the detection of methylated DNA site. Rolling circle amplification and CdSe/ZnS quantum dots with high fluorescence efficiency are applied in this method. The CdSe/ZnS quantum dots act as electron donors, and hemin and oxygen (derived from hydrogen peroxide act as acceptors in photoinduced electron transfer. The assay, best performed at excitation/emission peaks of 450/620 nm, is sensitive and specific. Fluorometric response is linear in the 1 pM to 100 nM DNA concentration range, and the lowest detectable concentration of methylated DNA is 142 fM (S/N =?3). The method is capable of recognizing 0.01% methylated DNA in a mixture of methylated/unmethylated DNA.
Graphical abstract A novel method for methylated sites detection in DNA is established. Rolling circle amplification and photoinduced electron transfer. CdSe/ZnS quantum dots with high fluorescence efficiency act as the electron donor, while G-quadruplex/hemin and hydrogen peroxide derived oxygen act as electron acceptor. It presents a linear response towards 1 pM to 100 nM methylated DNA with a correlation coefficient of 0.9968, and the lowest detectable concentration of methylated DNA was 142 fM, with selectivity significantly superior to other methods.
The authors describe an aptamer based assay for determination of ractopamine (RAC) by using PicoGreen (PG) as a fluorescent probe specific for dsDNA. In the absence of RAC, the aptamer forms a duplex structure with a complementary sequence that results in enhanced PG fluorescence. Upon binding to RAC, the aptamer undergoes a structural switch. This reduces the number of DNA duplexes formed and causes a reduction of fluorescence intensity of PG as measured at excitation/emission wavelengths of 480/520 nm. Under optimized conditions, the dynamic calibration plot covers the 50 pM to 50 μM concentration range, with a 50 pM detection limit. This meets the safety supervision regulations of the European Commission in terms of residue limits of RAC in food. The method displays high selectivity over other β-adrenergic agonists including clenbuterol, dopamine and salbutamol. The assay was successfully applied to samples of swine urine at spiking levels of 7.4 nM, 22.2 nM and 37 nM. Average recoveries ranged from 95 to 110%, with an RSD of <1.5%. The method is expected to represent a promising tool for simple, rapid and sensitive on-site detection of RAC in animal products.
Graphical abstract An aptamer based fluorescent assay for determination of ractopamine was developed with a dynamic range of 50 pM to 50 μM. The average recovery from spiked urine samples ranged from 95 to 110%, with an RSD of <1.5%.
This paper describes a CdTe quantum dot-based fluorescence resonance energy transfer (FRET) based assay for the detection of the breast cancer biomarker microRNA. The method relies on energy transfer between DNA-templated silver nanoclusters (AgNCs) and CdTe QDs. Interaction between double strand oligonucleotide and QDs can be detected qualitatively through gel analysis and quantitatively by the signal amplification from AgNCs to QDs via FRET, best measured at an excitation wavelength of 350 nm and at emission wavelengths of 550 and 590 nm. Three microRNAs (microRNA-21, microRNA-155 and Let-7a) were quantified to verify the feasibility of the method, and a high sensitivity for microRNAs was achieved. Fluorescence intensity increases linearly with the log of the concentration of microRNA 155 in the 5.0 pM to 50 nM range, with a 1.2 pM detection limit.
Graphical abstract Schematic presentation of a quantum dot-based (QD-based) fluorescence resonance energy transfer technique for the detection of microRNA (miRNA). The method relies on energy transfer between DNA-templated silver nanoclusters (AgNCs) and QDs.
The work describes a hybrid electrochemical sensor for highly sensitive detection of the anesthetic lidocaine (LID). Porous carbon (PC) was synthesized from an isoreticular metal-organic framework-8 (IRMOF-8) and drop cast onto a glassy carbon electrode (GCE). A layer of a molecularly imprinted polymer (MIP) layer was then fabricated in situ on the modified GCE by electro-polymerization, with LID acting as the template and resorcinol as the functional monomer. Hexacyanoferrate is used as an electrochemical probe. The electrical signal (typically acquired at 0.335 V vs. SCE) increases linearly in the 0.2 pM to 8 nM LID concentration range, with a remarkable 67 fM detection limit (at an S/N ratio of 3). The sensor is stable and selective. Eventually, rapid and accurate detection of LID in spiked real samples was successfully realized.
A fluorometric ATP assay is described that makes use of carbon dots and graphene oxide along with toehold-mediated strand displacement reaction. In the absence of target, the fluorescence of carbon dots (with excitation/emission maxima at 360/447 nm) is strong and in the “on” state, because the signal probe hybridizes with the aptamer strand and cannot combine with graphene oxide. In the presence of ATP, it will bind to the aptamer and induce a strand displacement reaction. Consequently, the signal probe is released, the sensing strategy will change into the “off” state with the addition of graphene oxide. This aptasensor exhibits selective and sensitive response to ATP and has a 3.3 nM detection limit.
Graphical abstract Schematic of signal amplification by strand displacement in a carbon dot based fluorometric assay for ATP. This strategy exhibits high sensitivity and selectivity with a detection limit as low as 3.3 nM.
Gold-silver nanoclusters (Au-AgNCs) were synthesized by simultaneous chemical reduction of Au(III) and Ag(I) ions in one pot, using bovine serum albumin as both a template and a reductant. The Au-AgNCs have an average size of 2.4 nm and display strong red fluorescence (with an emission peak at 610 nm on excitation at 360 nm). The fluorescence quantum yield can reach 18.6%. Fluorescence is strongly quenched by hypochlorite, while other common anions have minor (or no) effects on fluorescence. Based on these findings, a fluorometric method was developed for the determination of hypochlorite. The method has a linear response in the 0.7 to 15 μM concentration range, with a limit of detection as low as 80 nM. It was successfully applied to the determination of hypochlorite in (spiked) tap water.
Graphical abstract Gold-silver nanoclusters with strong red fluorescence were synthesized by simultaneous chemical reduction of Au(III) and Ag(I) ions in one pot, and a sensitive and selective method for the detection of hypochlorite was developed based on the quenching of the fluorescence of the nanoclusters.
The authors report on a one-pot approach for synthesizing highly fluorescent protamine-stabilized gold nanoclusters. These are shown to be a viable nanoprobe for selective and sensitive fluorometric determination of lead(II) via quenching of fluorescence via Pb(II)-Au(I) interaction. Under optimized conditions, fluorescence measured at excitation/emission peaks of 300/599 nm drops in the 80 nM–15 μM lead(II) concentration range. The detection limit is 24 nM, and relative standard deviations (for n?=?11) at concentrations of 0.10, 4.0 and 15 μM are 1.6, 2.5 and 1.9%, respectively. The relative recoveries of added lead(II) in the water samples ranged from 97.9?±?2.29% to 101.2?±?1.83%.
Graphical abstract Lead(II) ions are found to be able to selectively and sensitively quench the fluorescence of the protamine-gold nanoclusters (PRT-AuNCs). Thereby, an inexpensive, selective and sensitive lead(II) assay was established.
A facile, one-pot green method is presented for the preparation of water-soluble luminescent copper nanoclusters (Cu-NCs) from copper dichloride and cysteine as the precursor and stabilizer, respectively. The Cu-NCs are characterized by high resolution transmission electron microscopy, X-ray photoelectron spectroscopy, fluorescence, UV–Vis, and Raman spectroscopy. The Cu-NCs have an average size of 3.5 nm and are stable in aqueous solution at least for 2 weeks. Under photo excitation with 365 nm light, the Cu-NCs display strong green fluorescence with the maximum of emission at 490 nm and a quantum yield of 5.6 %. Fluorescence is quenched by Cr(VI) ion, and this effect was exploited to develop a highly selective method for the determination of Cr(VI). The detection limit of this probe is as low as 43 nM.
Graphical Abstract A facile, one-pot, “green” synthetic route was developed for preparing water-soluble luminescent copper nanoclusters (CuNCs) by using copper chloride and cysteine as the precursor and stabilizer, respectively. Their fluorescence is quenched by Cr(VI) ion, and this is exploited in a sensitive assay for Cr(VI) ions.
An electrochemical microsensor for chloramphenicol (CAP) was fabricated by introducing magnetic Fe3O4 nanoparticles (NPs) onto the surface of activated carbon fibers. This microsensor exhibited increased electrochemical response toward CAP because of the synergetic effect of the Fe3O4 NPs and the carbon fibers. Cyclic voltammograms were acquired and displayed three stable and irreversible redox peaks in pH 7.0 solution. Under optimized conditions, the cathodic current peaks at ?0.67 V (vs. Ag/AgCl). The calibration plot is linear in the 40 pM to 1 μM CAP concentration range, with a 17 pM detection limit (at a signal-to-noise ratio of 3). The sensor was applied to the determination of CAP in spiked sediment samples. In our perception, this electrocatalytic platform provided a useful tool for fast, portable, and sensitive analysis of chloramphenicol.
Graphical abstract A sensitive carbon fiber microsensor modified with Fe3O4 nanoparticles is found to display two cathodic peaks when detecting chloramphenicol at 100 mV·s?1 and at pH 7.0. The sensor was applied to the determination of chloramphenicol in sediment samples.
The authors describe an electrochemical sensing strategy for highly sensitive and specific detection of target (analyte) DNA based on an amplification scheme mediated by a multicomponent nucleic acid enzyme (MNAzyme). MNAzymes were formed by multicomponent complexes which produce amplified “output” signals in response to specific “input” signal. In the presence of target nucleic acid, multiple partial enzymes (partzymes) oligonucleotides are assembled to form active MNAzymes. These can cleave H0 substrate into two pieces, thereby releasing the activated MNAzyme to undergo an additional cycle of amplification. Here, the two pieces contain a biotin-tagged sequence and a byproduct. The biotin-tagged sequences are specifically captured by the detection probes immobilized on the gold electrode. By employing streptavidinylated alkaline phosphatase as an enzyme label, an electrochemical signal is obtained. The electrode, if operated at a working potential of 0.25 V (vs. Ag/AgCl) in solution of pH 7.5, covers the 100 pM to 0.25 μM DNA concentration range, with a 79 pM detection limit. In our perception, the strategy introduced here has a wider potential in that it may be applied to molecular diagnostics and pathogen detection.
Graphical abstract An electrochemical strategy for sequence-specific DNA detection based on multicomponent nucleic acid enzyme (MNAzyme) -mediated signal amplification.
Copper nanoclusters (Cu-NCs) were prepared by reducing CuCl2 with ascorbic acid in the presence of the short peptide template Cys-Cys-Cys-Asp-Leu. They were characterized by UV-vis absorption and fluorescence spectroscopy, transmission electron microscopy and X-ray photoelectron spectroscopy. The Cu-NCs have a size of ~2 nm, can be well dispersed in water and are photostable. Their fluorescence (peaking at 425 nm under 365-nm excitation) is quenched by Fe(III) ions. Based on this finding, a sensitive and selective fluorescence assay for the detection of Fe(III) was developed. Under optimized conditions and a pH value of 2.0, the assay displays a linear response in the 0.05 to 30 μM Fe(III) concentration range, with a detection limit of 20 nM based on an S/N ratio of 3. The assay was successfully applied to the determination of Fe(III) in spiked human serum where is gave recoveries that ranged from 96.2 % to 98.3 %.
Graphical abstract Copper nanoclusters (Cu-NCs) were prepared by reducing CuCl2 with ascorbic acid with peptide as the template. The fluorescence of Cu-NCs is quenched by Fe(III) ions with a linear response in the 0.05 to 30 μM of Fe(III) concentration range.
CdTe quantum dots (QDs) were integrated with polyethyleneimine-coated carbon dots (PEI-CDs) to form a dually emitting probe for heparin. The red fluorescence of the CdTe QDs is quenched by the PEI-CDs due to electrostatic interactions. In the presence of heparin, the blue fluorescence of PEI-CDs remains unaffected, while its quenching effect on the fluorescence of CdTe QDs is strongly reduced. A ratiometric fluorometric assay was worked out. The ratio of the fluorescences at 595 and 436 nm serves as the analytical signal. Response is linear in the concentration range of 50–600 ng·mL?1 (0.1–1.2 U·mL?1) of heparin. The limit of detection is 20 ng·mL?1 (0.04 U·mL?1). This makes the method a valuable tool for heparin monitoring during postoperative and long-term care. This assay is relatively free from the interference by other analogues which commonly co-exist with heparin in samples, and it is more robust than single-wavelength based assays.
Graphical abstract In the presence of heparin, the fluorescence of polyethyleneimine-coated carbon dots (PEI-CDs) at 436 nm remains unaffected, while its quenching effect on the fluorescence of CdTe at 595 nm is strongly reduced.
It is found that the fluorescence of carbon dots (CD) with an emission peak at 459 nm is strongly quenched by silver nanoparticles (AgNPs) with their absorption peak at 430 nm. The finding was applied in a fluorescence quenchometric lateral flow immunochromatographic assay for detection of zearalenone (ZEN) with CDs conjugated to ovalbumin (OVA) as donor signal probe and AgNP-Ab as acceptor signal probe. The assay has an LOD of 0.1 μg·L?1 for ZEN. This is 10 times better than the respective “turn-off” AgNP-based LFIA. In case of cereal samples and their products, the LODs range from 1 to 2.5 μg·kg?1. Only minor cross reactivity is found for fusarium toxins, and no cross-sensitivity for aflatoxin B1, T-2 mycotoxin, ochratoxin A, deoxynivalenol, and fumonisin B1. The assay represents a simple, sensitive, and rapid tool for determination of ZEN in cereal samples and their products.
Graphical abstract Schematic presentation of fluorescence quenching lateral flow immunochromatographic assay (FLFIA) based on carbon dots (CD) and silver nanoparticle (AgNP) fluorescence resonance energy transfer (FRET) system for the rapid high sensitive detection of zearalenone (ZEN) in cereal samples.
The authors describe a fluorescence based aptasensor for adenosine (AD), a conceivable biomarker for cancer. The assay is based on the immobilization of capture DNA on newly synthesized quaternary CuInZnS quantum dots (QDs) and the conjugation of probe DNA on gold nanoparticles (AuNPs). The capture DNA is an adenosine-specific aptamer that is partly complementary to the probe DNA. Once the capture aptamer hybridizes probe DNA, the fluorescence of the QDs (measured at excitation/emission wavelengths of 522/650 nm) is quenched by the AuNPs. However, when AD is added, it will bind to the aptamer and restrain the hybridization between capture DNA and probe DNA. Therefore, the fluorescence of the QDs will increase with increasing AD concentration. Under optimal conditions, fluorescence is linearly related to the AD concentration in the range from 50 to 400 μM, the detection limit being 1.1 μM. This assay is sensitive, selective, reproducible and acceptably stable. It was applied to the determination of AD in spiked human serum samples where it gave satisfactory results.
Hydroxyapatite nanoparticles (HAP-NPs) were rendered fluorescence by doping with Eu(III) ion. The resulting fluorescent NPs are shown to be viable probes for sensitive and selective determination of dipicolinic acid (DPA), a major constituent of bacterial spores as used in bioterrorism. It is found that the addition of DPA to solutions of such HAP-NPs result in an enhancement of fluorescence due to the coordination of DPA with the Eu(III) dopant. The assay allows DPA to be detected in the 0.1 to 40 μM concentration range and with a 77 nM detection limit. The assay was applied to the detection of spores of Bacillus subtilis. The attractive properties of the probe make it a promising candidate for used in rapid detection of pathogenic bacterial spores.
Graphical abstract Fluorescent hydroxyapatite nanoparticles (HAP-NPs) are shown to be a viable probe for detection of dipicolinic acid, a major constituent of bacterial spores. The red asterisks represent the fluorescence intensity of the HAP-NPs.
A method is described for the determination of the activity of alkaline phosphatase (ALP). It is based on the reversible modulation of the fluorescence of WS2 quantum dots (QDs). The fluorescence of the QDs is quenched by Cr(VI) but restored by free ascorbic acid (AA). The detection scheme relies on the fact that ALP hydrolyzes the substrate ascorbic acid 2-phosphate to produce AA, and that enzymatically generated AA can restore the fluorescence of the QDs. The signal (best measured at excitation/emission peak wavelengths of 365/440 nm) increases linearly in the 0.5 to 10 U·L?1 ALP activity range, with a detection limit of 0.2 U·L?1. The method was applied to the determination of ALP activity in human serum samples and demonstrated satisfactory results.
Graphical abstract The fluorescence of chromate-loaded tungsten disulfide quantum dots (QDs) is quenched but restored after reaction with ascorbic acid that is formed by the catalytic action of alkaline phosphatase (ALP) on ascorbic acid 2-phosphate (AAP). The increase in fluorescence can be related to the activity of ALP.
Europium(III)-doped carbon dots (Eu-CDs) were prepared from citric acid and europium nitrate via a one-pot pyrolytic method. The Eu-CDs emit intense blue fluorescence (with excitation/emission peaks at 365/465 nm), are water soluble and biocompatible. On addition of 2,6-dipicolinic acid (DPA; an anthrax biomarker), ligand-to-ion energy transfer occurs from DPA to Eu(III) which has a red emission peaking at 615 nm. This results in an increase of the intensity of the red fluorescence. DPA can be detected by the ratio of fluorescence intensities at 616 and 475 nm. The method has an analytical range that extends from 5 to 700 nmol·L?1, with a 5 nmol·L?1 detection limit. The Eu-CDs also were incorporated into a test paper for visual detection of DPA with a portable UV lamp and a smartphone. In this case, the detection limit is 1 μmol·L?1. The Eu-CDs internalize well into HeLa cells, and this paves the way to bioimaging.
Graphical abstract Schematic of a method for visual detection of 2,6-dipicolinic acid (DPA, an anthrax biomarker) by using a test stripe impregnated with europium(III)-doped carbon dots (Eu-CDs).
Cobalt oxyhydroxide (CoOOH) nanosheets are efficient fluorescence quenchers due to their specific optical properties and high surface area. The combination of CoOOH nanosheets and carbon dots (CDs) has not been used in any aptasensor based on fluorescence quenching so far. An aptamer based fluorometric assay is introduced that is making use of fluorescent CDs conjugated to the aptamer against methamphetamine (MTA), and of CoOOH nanosheets which reduce the fluorescence of the CDs as a quencher. The results revealed that the conjugated CDs with aptamers were able to enclose the CoOOH nanosheets. Consequently, fluorescence is quenched. If the aptamer on the CD binds MTA, the CDs are detached from CoOOH nanosheets. As a result, fluorescence is restored proportionally to zhe MTA concentration. The fluorometric limit of detection is 1 nM with a dynamic range from 5 to 156 nM. The method was validated by comparing the results obtained by the new method to those obtained by ion mobility spectroscopy. Theoretical studies showed that the distance between CoOOH nanosheet and C-Ds is approximately 7.6 Å which can illustrate the possibility of FRET phenomenon. The interactions of MTA and the aptamer were investigated using molecular dynamic simulation (MDS).
Graphical abstract Carbon dots (C-Ds) were prepared from grape leaves, conjugated to aptamer, and adsorbed on CoOOH nanosheets. So, the fluorescence of C-Ds is quenched. On addition of MTA, fluorescence is restored.
