基于胸腺嘧啶-汞离子-胸腺嘧啶结构和纳米金放大传感器检测汞离子

利用Hg2+与DNA中胸腺嘧啶(T)结合的高度特异性和纳米金在石英晶体微天平(QCM)上的信号放大作用,设计了一种简便灵敏的Hg2+检测方法.纳米金采用柠檬酸钠还原法制备,其表面用末端带巯基的寡核苷酸探针进行自组装修饰,并用6-巯基己-1-醇(MCH)部分取代表面探针,以减少杂交空间位阻.结果表明,寡核苷酸链长为9bp、T个数为7的序列具有较高灵敏度;线性范围为5.0～100 nmol/L;检出限为2.0 nmol/L;Ca2+、Mg2+等其它金属离子无明显干扰.用于环境水样中Hg2+的测定, RSD<2.9%;加标回收率为97.3%～101.2%     

Colorimetric assay for mercury (II) based on mercury-specific deoxyribonucleic acid-functionalized gold nanoparticles

A colorimetric nanoprobe-mercury-specific DNA-functionalized gold nanoparticles (Au-MSD) was developed for sensing Hg(2+). The new mercury-sensing concept relies on measuring changes in the inhibition of "non-crosslinking" aggregation of Au-MSD-induced by the folding of mercury-specific DNA strand through the thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination. In the absence of Hg(2+), a high concentration of MgCl(2) (50 mM) results in a rapid aggregation of Au-MSD because of the removal of charge repulsion. When Hg(2+) is present, the particles remain stable due to the folding of MSD functionalized on the particle surface. The assay enables the colorimetric detection of Hg(2+) in the concentration range of 0.1-10 μM Hg(2+) ions with a detection limit of 60 nM, and allows for the selective discrimination of Hg(2+) ions from the other competitive metal ions. Toward the goal for practical applications, the sensor was further evaluated by monitoring Hg(2+) in fish tissue samples.     

Oligonucleotide-functionalized gold nanoparticles-enhanced QCM-D sensor for mercury(II) ions with high sensitivity and tunable dynamic range

A quartz crystal microbalance with dissipation monitoring (QCM-D) sensor was developed for highly sensitive and specific detection of mercury(II) ions (Hg(2+)) with a tunable dynamic range, using oligonucleotide-functionalized gold nanoparticles (GNPs) for both frequency and dissipation amplification. The fabrication of the sensor employed a 'sandwich-type' strategy, and formation of T-Hg(2+)-T structures in linker DNA reduced the hybridization of the GNPs-tagged DNA on the gold electrode, which could be used as the molecular switch for Hg(2+) sensing. This QCM-D mercury sensor showed a linear response of 10-200 nM, with detection limits of 4 nM and 7 nM for frequency and dissipation measurements, respectively. Moreover, the dynamic range of the sensor could be tuned by simply altering the concentration of linker DNA without designing new sensors in the cases where detection of Hg(2+) at different levels is required. This sensor afforded excellent selectivity toward Hg(2+) compared with other potential coexisting metal ions. The feasibility of the sensor was demonstrated by analyzing Hg(2+)-spiked tap- and lake-water samples with satisfactory recoveries. The proposed approach extended the application of the QCM-D system in metal ions sensing, and could be adopted for the detection of other analytes when complemented with the use of functional DNA structures.     

Highly sensitive, selective, and rapid fluorescence Hg2+ sensor based on DNA duplexes of poly(dT) and graphene oxide

Coupling T base with Hg(2+) to form stable T-Hg(2+)-T complexes represents a new direction in detection of Hg(2+). Here a graphene oxide (GO)-based fluorescence Hg(2+) analysis using DNA duplexes of poly(dT) that allows rapid, sensitive, and selective detection is first reported. The Hg(2+)-induced T(15)-(Hg(2+))(n)-T(15) duplexes make T(15) unable to hybridize with its complementary A(15) labelled with 6'-carboxyfluorescein (FAM-A(15)), which has low fluorescence in the presence of GO. On the contrary, when T(15) hybridizes with FAM-A(15) to form double-stranded DNA because of the absence of Hg(2+), the fluorescence largely remains in the presence of GO. A linear range from 10 nM to 2.0 μM (R(2) = 0.9963) and a detection limit of 0.5 nM for Hg(2+) were obtained under optimal experimental conditions. Other metal ions, such as Al(3+), Ag(+), Ca(2+), Ba(2+), Mg(2+), Zn(2+), Mn(2+), Co(2+), Pb(2+), Ni(2+), Cu(2+), Cd(2+), Cr(3+), Fe(2+), and Fe(3+), had no significant effect on Hg(2+) detection. Moreover, the sensing system was used for the determination of Hg(2+) in river water samples with satisfactory results.     

镊子型dsDNA稳定的纳米金光度法快速检测Hg2+的研究

利用Hg2+对胸腺嘧啶(T)T -T错配的特异性结合,建立了一种利用盐诱导金纳米粒子聚集的比色定量检测Hg2+离子的方法.设计了一种镊子型dsDNA,其一半为互补碱基形成的双螺旋结构,另一半为T-T错配.错配部分保持单链状态吸附在纳米金表面,使纳米金的稳定性增强,抑制盐诱导的纳米金团聚.当存在Hg2时,“T- Hg2+...     

Amplified fluorescence detection of mercury(II) ions (Hg2+) using target-induced DNAzyme cascade with catalytic and molecular beacons

In this work, a fluorescent sensing strategy was developed for the detection of mercury(II) ions (Hg(2+)) in aqueous solution with excellent sensitivity and selectivity using a target-induced DNAzyme cascade with catalytic and molecular beacons (CAMB). In order to construct the biosensor, a Mg(2+)-dependent DNAzyme was elaborately designed and artificially split into two separate oligonucleotide fragments. In the presence of Hg(2+), the specific thymine-Hg(2+)-thymine (T-Hg(2+)-T) interaction induced the two fragments to produce the activated Mg(2+)-dependent DNAzyme, which would hybridize with a hairpin-structured MB substrate to form the CAMB system. Eventually, each target-induced activated DNAzyme could catalyze the cleavage of many MB substrates through true enzymatic multiple turnovers. This would significantly enhance the sensitivity of the Hg(2+) sensing system and push the detection limit down to 0.2 nM within a 20 min assay time, much lower than those of most previously reported fluorescence assays. Owning to the strong coordination of Hg(2+) to the T-T mismatched pairs, this proposed sensing system exhibited excellent selectivity for Hg(2+) detection, even in the presence of 100 times of other interferential metal ions. Furthermore, the applicability of the biosensor for Hg(2+) detection in river water samples was demonstrated with satisfactory results. These advantages endow the sensing strategy with a great potential for the simple, rapid, sensitive, and specific detection of Hg(2+) from a wide range of real samples.     

Visual and on-site detection of mercury(II) ions on lateral flow strips using DNA-functionalized gold nanoparticles

A test strip for detection of Hg(2+) in aqueous solution based on the DNA-functionalized gold nanoparticles (DNA-AuNPs) was developed and evaluated. When Hg(2+) ions were introduced, the biotinylated DNA(2) hybridized with thiolated DNA(1) functionalized on the AuNPs (DNA(1)-AuNPs) to form mismatch complexes through thymine-Hg(2+)-thymine (T-Hg(2+)-T) coordination. The formed mismatch complexes and excess DNA(1)-AuNPs could be captured on the test line formed by streptavidin and the control line formed by DNA(3)-BSA, respectively. Two red lines appeared due to the accumulation of AuNPs, enabling visual detection of Hg(2+) with a detection limit of about 6 nM. The assay results can be obtained within 5 min. The results show that the test strip has excellent sensitivity and selectivity for detection of Hg(2+); thus it holds a great potential for rapid, on-site and real time detection of Hg(2+).     

A simple and sensitive fluorescent sensing platform for Hg²+ ions assay based on G-quenching

In this work, a novel fluorescence biosensor was demonstrated for detection of Hg(2+) ions with relatively high selectivity and sensitivity. The sensing scheme was based on G-quenching induced by Hg(2+) ions. In the presence of Hg(2+) ions, the single-stranded signal probe which has carboxylfluorescein (FAM) and guanine segment at its 5' and 3' ends, respectively, folded into duplex-like structure via the Hg(2+)-mediated coordination of T-Hg(2+)-T base pairs. It brought guannine segment close to the dye and caused a remarkable decrease of fluorescence signal. The sensor showed a sensitive response to Hg(2+) ions in a concentration range from 0.5 to 10 μM, and a detection limit of 0.5 nM was given. This homogeneous system required only a single-labeled oligonucleotide, operated by concise procedures, and possessed comparable sensitivity as previous approaches. Furthermore, the sensor exhibits a great perspective for future practical applications.     

Logic gates for multiplexed analysis of Hg2+ and Ag+

The design of devices with multiple functions, simple handling procedures and sufficient sensitivity has drawn great interests in the field of analysis. Metal-nucleotide based pairs, such as T-Hg(2+)-T and C-Ag(+)-C complexes accompanied by SYBR Green I (SG), are used to selectively bind duplex-strand DNA by observing a bright fluorescence signal in this work, thus yielding a simple method for the rapid detection of Hg(2+) and Ag(+) without a complex labeling process. Based on this principle, 'OR' and 'AND' logic gates for the multiplexed analysis of Hg(2+) and Ag(+) were developed, and their practical application for the detection of Hg(2+) and Ag(+) in drinking water was reported.     

Gold nanoparticles generated through "green route" bind Hg2+ with a concomitant blue shift in plasmon absorption peak

We discuss here a quick, simple, economic and ecofriendly method through a completely green route for the selective detection of Hg(2+) in aqueous samples. Here we exploited the ability of chitosan to generate gold nanoparticles and subsequently to act as a stabilizer for the formed nanoparticles. When chitosan stabilized gold nanoparticles (CH-Au NPs) are interacted with Hg(2+) a blue shift for its localized surface plasmon resonance absorbance (LSPR) band is observed. The blue shift is reasoned to be due to the formation of a thin layer of mercury over gold. A concentration as low as 0.01 ppm to a maximum of 100 ppm Hg(2+) can be detected based on this blue shift of the CH-Au NPs. While all other reported methods demand complex reaction steps and costly chemicals, the method we reported here is a simple, rapid and selective approach for the detection of Hg(2+). Our results also show that the CH-Au NPs have excellent selectivity to Hg(2+) over common cations namely, Pb(2+), Cd(2+), Mn(2+), Fe(2+), Ag(1+), Ce(4+), Ni(2+), and Cu(2+).     

基于纳米金Core-satellites等离子体耦合增强效应的汞离子光纤传感器的研究

以DNA杂交双链为联接, 构建纳米金颗粒Core-satellites结构并激发等离子体耦合增强效应,利用Hg2+可与DNA中胸腺嘧啶T形成T-Hg2+-T特异性结构,研制了用于检测水中Hg2+的局域等离子体共振(LSPR)光纤传感器.待测溶液中的Hg2+能够引起富含T的DNA单链折叠,抑制DNA杂交反应,降低等离子体耦合强度,改变LSPR谐振波长.通过检测谐振波长红移变化,实现对Hg2+浓度的定量检测.本方法检测Hg2+的线性范围为5~150 nmol/L, 检出限为3.4 nmol/L (3σ). Zn2+、Mg2+、Pb2+等重金属离子对Hg2+检测无明显干扰作用.实际水样中Hg2+加样回收率为94.2%~105.4%,相对标准偏差<4.8%.     

A one-step colorimetric method of analysis detection of Hg2+ based on an in situ formation of Au@HgS core-shell structures

A new approach for the detection of Hg(2+) is reported based on color changes from which gold nanoparticles (Au NPs) are surrounded by a layer of HgS quantum dots to form in situ Au@HgS core-shell nanostructures. The surface plasmon resonance (SPR) absorption of the gold core was changed due to a shell layer of HgS formed on the surface of the Au NPs, which brings the colour change of the aqueous solution. Therefore, Hg(2+) can be recognized by visualizing the colour change of the Au@HgS core-shell nanostructures, and can be detected quantitatively by measurement of the UV-vis spectra. Some effects on the detection of Hg(2+) were investigated in detail. This method was used to detect Hg(2+) with excellent selectivity and high sensitivity. In our method, the lowest detected concentrations for mercury ions were 5.0 × 10(-6) M observed by the naked eye and 0.486 nM as measured by UV-vis spectra. At the range from 8.0 × 10(-5) to 1.0 × 10(-8) M of Hg(2+), this method was shown to have a good linear relationship.     

Label-free emission assay of mercuric ions using DNA duplexes of poly(dT)

In this study, an assay to quantify the presence of mercuric ions and methyl mercury by double-stranded DNA containing a poly(dT) sequence was developed using a light switch compound, Ru(phen)(2)(dppz)(2+) (1), which is known to intercalate into double-stranded DNA. Upon treatment with mercuric ions, the metal-to-ligand charge transfer (MLCT) emission derived from the intercalation of 1 was reduced due to the formation of DNA duplexes containing T-Hg(2+)-T base pairs by the dehybridization of poly(dT)-poly(dA) duplexes at room temperature. As the concentration of Hg(2+) was increased, the emission of 1 gradually decreased. This label-free method had a detection limit of 5 nM. Other metal ions, such as K(+), Ag(+), Ca(2+), Mg(2+), Zn(2+), Mn(2+), Co(2+), Ni(2+), Cu(2+), Cd(2+), Cr(3+), Fe(3+), had no significant effect on reducing emission. This emission method can differentiate matched and mismatched poly(dT) sequences based on the dehybridization rate of dsDNA and the rate decreased in the order of T(10)C·A(11)～ T(10)A·A(11) > T(10)G·A(11) > T(11)·A(11).     

Control over surface DNA density on gold nanoparticles allows selective and sensitive detection of mercury(II)

We have developed a new highly selective and sensitive technique for the detection of Hg(2+) using DNA-functionalized gold nanoparticles (Au NPs) and OliGreen. This system is the first that allows the detection of Hg(2+) based on the release of DNA molecules, induced by conformational changes on Au NP surfaces, and its sensitivity is highly dependent upon surface DNA density. When Hg(2+) ions interact with the thymidine units of the DNA molecules bound to the Au NPs through Au-S bonds, the conformations of these DNA derivatives change from linear to hairpin structures, causing the release of some of the DNA molecules from the surface of the Au NPs into the bulk solution to react with OliGreen. The fluorescence of OliGreen-DNA complexes increased with increasing concentration of Hg(2+), and Hg(2+) could be detected at concentrations as low as 25 nM. A linear correlation existed between the fluorescence intensity and the concentration of Hg(2+) over the range 0.05-2.5 microM (R(2) = 0.98). This simple and cost-effective probe was applied to determine the spiked Hg(2+) in the pond samples; the recoveries (96-102%) suggested low matrix interference and high sensitivity.     

An ultrasensitive electrochemical sensor for the mercuric ion via controlled assembly of SWCNTs

An ultrasensitive "turn-on" electrochemical sensor for the Hg(2+) ion was proposed based on the T-Hg(2+)-T coordination chemistry and the controlled assembly of SWCNTs on the MHA/SAM-modified gold electrode.     

Multiplexed analysis of silver(I) and mercury(II) ions using oligonucletide-metal nanoparticle conjugates

A colorimetric assay has been developed for the simultaneous selective detection of silver(I) and mercury(II) ions utilizing metal nanoparticles (NPs) as sensing element based on their unique surface plasmon resonance properties. In this method, sulfhydryl group modified cytosine-(C)-rich ssDNA (SH-C-ssDNA) was self-assembled on gold nanoparticles (AuNPs) to produce the AuNPs-C-ssDNA complex, and sulfhydryl group modified thymine-(T)-rich ssDNA (SH-T-ssDNA) was self-assembled on silver nanoparticles (AgNPs) to produce the AgNPs-T-ssDNA complex. Oligonucleotides (SH-C-ssDNA or SH-T-ssDNA) could enhance the AuNPs or AgNPs against salt-induced aggregation. However, the presence of silver(I) ions (Ag(+)) in the complex of ssDNA-AuNPs would reduce the stability of AuNPs due to the formation of Ag(+) mediated C-Ag(+)-C base pairs accompanied with the AuNPs color change from red to purple or even to dark blue. Moreover, the presence of mercury(II) ions (Hg(2+)) would also reduce the stability of AgNPs due to the formation of Hg(2+) mediated T-Hg(2+)-T base pairs accompanied with the AgNPs color change from yellow to brown, then to dark purple. The presence of both Ag(+) and Hg(2+) will reduce the stability of both AuNPs and AgNPs and cause the visible color change. As a result, Ag(+) and Hg(2+) could be detected qualitatively and quantitatively by the naked eye or by UV-vis spectral measurement. The lowest detectable concentration of a 5 nM mixture of Ag(+) and Hg(2+) in the river water was gotten by the UV-vis spectral measurement.     

Label-free supersandwich electrochemiluminescence assay for detection of sub-nanomolar Hg2+

A label-free supersandwich electrochemiluminescence assay based on T-Hg(2+)-T coordination and the intercalation of Ru(phen)(3)(2+) for Hg(2+) analysis was developed with excellent sensitivity and selectivity.     

N-1-(2-mercaptoethyl)thymine modification of gold nanoparticles: a highly selective and sensitive colorimetric chemosensor for Hg2+

An approach for mercury ions (Hg(2+)) sensing based on the Hg(2+)-induced aggregation of thymine (T)-SH-functionalized gold nanoparticles (AuNPs) has been reported. The T-SH ligands that we synthesized can easily be coupled to the surface of AuNPs through the Au-S bond and can recognize Hg(2+) with high selectivity by forming a T-Hg-T complex with strong affinity. For the T-SH-functionalized AuNPs (T-S-AuNPs) sensor, upon addition of Hg(2+), the formation of the T-Hg-T complex induces aggregation of T-S-AuNPs and results in a significant change of color and UV-Vis absorption spectra. Thus, our method can be used for the rapid, easy and reliable screening of Hg(2+) in aqueous solution, with high sensitivity (2.8 nM) and selectivity over competing analytes. The developed method is successfully applied to the sensing of Hg(2+) in real environmental samples.     

Coordination of mercury(II) to gold nanoparticle associated nitrotriazole towards sensitive colorimetric detection of mercuric ion with a tunable dynamic range

A simple colorimetric assay with high sensitivity, excellent selectivity and a tunable dynamic range is reported for detecting trace amounts of mercuric ion in aqueous solution based on the coordination of Hg(2+) to the gold nanoparticle (AuNP)-associated 3-nitro-1H-1,2,4-triazole (NTA). The NTA can stabilize the AuNPs against tris-induced aggregation through capping the AuNPs. In the presence of Hg(2+), the NTA is released from the AuNP surface via the formation of a NTA-Hg(2+) coordination complex, leading to the aggregation of AuNPs in tris. This detection strategy is unique in terms of high sensitivity and excellent selectivity, a tunable dynamic range, and simplicity of probe preparation. Low detection limits of 7 nM (1.4 ppb) and 50 nM (10 ppb) can be achieved by spectrophotometer and by direct visualization, respectively, under the optimized conditions. No noticeable colour changes are observed towards other metal ions (Ag(+), Zn(2+), Ni(2+), Cr(3+), Mg(2+), Cu(2+), Co(2+), Cd(2+), Pb(2+), Fe(2+)) at concentrations up to 100 μM without the need of any other masking agents. In addition, the dynamic range of the assay can be easily tuned by adjusting the amount of NTA in the NTA-AuNP probes. More importantly, the NTA-AuNP probes can be simply prepared by mixing NTA with as-synthesized citrate-capped AuNPs. This not only avoids complicated surface modifications and tedious separation processes, but also is cost-effective.     

Colorimetric detection of Fe3+ ions using pyrophosphate functionalized gold nanoparticles

A sensitive, selective colorimetric Fe(3+) detection method has been developed by using pyrophosphate functionalized gold nanoparticles (P(2)O(7)(4-)-AuNPs). Gold nanoparticles were prepared by reducing HAuCl(4) with sodium borohydride, in the presence of Na(4)P(2)O(7). IR spectra suggested that pyrophosphates were capped on the surface of the gold nanoparticles. Aggregation of P(2)O(7)(4-)-AuNPs was induced immediately in the presence of Fe(3+) ions, yielding a color change from pink to violet. This Fe(3+)-induced aggregation of P(2)O(7)(4-)-AuNPs was monitored using first the naked eye and then UV-vis spectroscopy with a detection limit of 5.6 μM. The P(2)O(7)(4-)-AuNPs bound by Fe(3+) showed excellent selectivity compared to other metal ions (Ca(2+), Cd(2+), Co(2+), Fe(2+), Hg(2+), K(+), Mg(2+), Mn(2+), Na(+), Ni(2+), Pb(2+), and Zn(2+)). The best detection of Fe(3+) was achieved in a pH range from 3 to 9. In addition, the P(2)O(7)(4-)-AuNPs were also used to detect Fe(3+) in lake water samples, with low interference.