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A novel fluorescence probe for the detection of Al3+ was developed based on methionine protected gold nanoclusters (Met-AuNCs). A fluorescent Schiff base (an aldimine) is formed between the aldehyde group of salicylaldehyde (SA) and the amino groups of Met on the AuNCs, and developed for selective detection of Al3+ in aqueous solution. Al3+ can strongly bind with the Schiff base ligands, accompanied by the blue-shift and an obvious fluorescence emission enhancement at 455 nm. The limits of detection (LODs) of the probe are 2 pmol L−1 for Al3+. Moreover, the probe can successfully be used in fluorescence imaging of Al3+ in living cells (SHSY5Y cells), suggesting that the simple fluorescent probe has great potential use in biological imaging. [ABSTRACT FROM AUTHOR]

A novelty aptasensor for ultrasensitive detection of Hg2+ is developed, exploiting the combination of plasmonic properties of gold nanoparticles (AuNPs) and exonuclease III (Exo III)-assisted target recycling for signal amplification. In the presence of Hg2+, a DNA duplex can be formed due to the strong coordination of Hg2+ and T bases of single-stranded DNA (ssDNA) probe. Exo III digests the DNA duplex from the 3′ to 5′ direction, resulting in the releasing of Hg2+. Then, the released Hg2+ binds with another ssDNA probe through T-Hg2+-T coordination. After Exo III-assisted Hg2+ cycles, numerous ssDNA probes are exhausted, which promotes poly(diallyldimethylammonium chloride) (PDDA)-induced AuNP aggregation, leading to an obvious color change and aggregation-induced plasmon red shift of AuNPs (from 520 to 610 nm). Therefore, this biosensor is ultrasensitive, which is applicable to the detection of trace level of Hg2+ with a linear range from 5 pM to 0.6 nM and an ultralow detection limit of 0.2 pM. Furthermore, it enables visual detection of Hg2+ as low as 50 pM by the naked eye. More importantly, the assay can be applied to the reliable determination of spiked Hg2+ in sea water samples with good recovery. [ABSTRACT FROM AUTHOR]

A facile plasmonic nanoplatform was developed for rapid and sensitive determination of nucleic acid. Hg2+-regulated molecular beacon (MB, hairpin) containing rich thymine (T) bases at both ends is used as the probe. A hairpin structure can be formed in the MB probe due to the strong binding of Hg2+ to T. However, in the presence of target DNA, the hairpin structure is opened owing to target DNA-specific hybridization with the aptamer. Simultaneously, the opened MB interacts with poly(diallyldimethylammonium chloride) (PDDA) and hinders PDDA-induced aggregation of AuNPs, accompanied by a color change from blue to red and a decrease in absorption ratio (A620/A520). Hence, a good linear relationship was observed between the decreased absorption ratio (A620/A520) and DNA concentration ranging from 0.02 to 2 nmol/L with a low detection limit of 4.42 pmol/L. Moreover, this nanoplatform has been successfully utilized to discriminate between perfect target and mismatch sequences. More importantly, the bioassay is simple, versatile, rapid, and cost-effective compared with other common methods, which holds great promise for clinical diagnosis and biomedical application. [ABSTRACT FROM AUTHOR]

A facile and rapid colorimetric approach was described for selective and sensitive determination of adenosine triphosphate (ATP) based on a hairpin aptamer probe and the anti-aggregation of AuNPs. Poly(diallyldimethylammonium chloride) (PDDA) can induce the aggregation of AuNPs due to the electrostatic interaction causing a red to blue color change. Upon the addition of ATP, aptamer-based hairpin probe is opened and releases flexible ssDNA ends. The released flexible ssDNA ends can interact with PDDA and prevent PDDA-induced AuNPs aggregation. Thus, a visible color change from blue to red and a decrease in the absorption ratio (A 610 /A 520) are observed. Under the optimal conditions, the hairpin aptamer-based colorimetric assay exhibits high sensibility and selectivity for the detection of ATP with a detection limit of 1.7 nM. Moreover, this assay is successfully used in the rapid determination of ATP in spiked human serum samples with good recoveries in the range of 102.88 to 104.07%. Schematic illustration of the colorimetric assay based on the anti-aggregation of AuNPs for the ATP detection. Flexible ssDNA ends were released due to the forming of the aptamer-ATP complexes, which electrostatically hybridize to PDDA to form a duplex and prevents the PDDA-induced aggregation of AuNPs with a color change from blue to red. Unlabelled Image • It has a high sensitivity with a detection limit of 1.7 nM for ATP. Furthermore, the coupling a hairpin-like DNA probe and anti-aggregation of AuNPs guarantees the good selectivity of the assay to ATP and eliminates the effect of non-specific interference. • It is a more universal method that can be used to study other targets by simply changing the sequences of the corresponding aptamers of the hairpin-like DNA probe. [ABSTRACT FROM AUTHOR]

In this work, a rapid, simple and sensitive colorimetric sensor for simultaneous (or respective) detection of Cr 3 + and Pb 2 + using tyrosine functionalized gold nanoparticles (AuNPs Tyr ) has been developed. Tyrosine, a natural and zwitterionic amino acid, could be as a reducing and capping agent to synthesise AuNPs and allow for the simultaneous and selective detection of Cr 3 + and Pb 2 + . Upon the addition of Cr 3 + or Pb 2 + (a combination of them), the color of AuNPs Tyr solution changes from red to blue grey and the characteristic surface plasmon resonance (SPR) band is red-shifted to 580 nm due to the aggregation of AuNPs. Interestingly, the aggregated AuNPs Tyr can be regnerated and recycled by removing Pb 2 + and Cr 3 + . Even after 3 rounds, AuNPs Tyr show almost the same A 580 nm / A 520 nm value for the assays of Pb 2 + and Cr 3 + , indicating the good recyclability of the colorimetric sensor. The responding time (within 1 min) and sensitivity of the colorimetric sensor are largely improved after the addition of 0.1 M NaCl. Moreover, the AuNPs Tyr aggregated by Cr 3 + or Pb 2 + (a combination of them) show excellent selectivity compared to other metal ions (Cr 3 + , Pb 2 + , Fe 2 + ,Cu 2 + ,Zn 2 + ,Cr 6 + ,Ni 2 + ,Co 2 + ,Hg 2 + ,Mn 2 + ,Mg 2 + ,Ca 2 + ,Cd 2 + ). More importantly, the developed sensor manifests good stability at room temperature for 3 months, which has been successfully used to determine Cr 3 + and Pb 2 + in the real water samples with a high sensitivity. [ABSTRACT FROM AUTHOR]

Multi-round PCR is an important technique for obtaining enough target DNA from rare DNA resources, and is commonly used in many fields including forensic science, ancient DNA analysis and cancer research. However, multi-round PCR is often aborted, largely due to the accumulation of non-specific amplification during repeated amplifications. Here, we developed a Pfu polymerase based multi-round PCR technique assisted by quantum dots (QDs). Different PCR assays, DNA polymerases (Pfu and Taq), DNA sizes and GC amounts were compared in this study. In the presence of QDs, PCR specificity could be retained even in the ninth-round amplification. Moreover, the longer and more complex the targets were, the earlier the abortion happened in multi-round PCR. However, no obvious enhancement of specificity was found in multi-round PCR using Taq DNA polymerase. Significantly, the fidelity of Pfu polymerase based multi-round PCR was not sacrificed in the presence of QDs. Besides, pre-incubation at 50 °C for an hour had no impact on multi-round PCR performance, which further authenticated the hot start effect of QDs modulated in multi-round PCR. The findings of this study demonstrated that a cost-effective and promising multi-round PCR technique for large-scale and high-throughput sample analysis could be established with high specificity, sensibility and accuracy. [ABSTRACT FROM AUTHOR]

Hot-start (HS) effects were investigated in pfu-based polymerase chain reaction (PCR), when water-soluble CdTe quantum dots (QDs) were introduced in the PCR system. The HS effects were demonstrated by the higher amplicon yields and excellent suppression of non-specific amplification after pre-incubation of PCR mix with QDs between 35°C and 56°C. DNA targets were well amplified even after PCR mixture was pre-incubated 1 h at 50°C. Importantly, the effects of QDs nanoparticles could be reversed by increasing the pfu polymerase concentration, suggesting that there was an interaction between QDs and pfu DNA polymerase. Moreover, control experiment indicated that HS effect is not primarily due to the reduced pfu polymerase concentration resulted from the above interaction. Fluorescence correlation spectroscopy (FCS), a single molecule detection method, was used to investigate the possible mechanism of HS PCR with QDs. Preliminary FCS results suggested that CdTe QDs may directly interact with pfu DNA polymerase, rather than other components in the PCR system. Furthermore, results demonstrated that the interaction between QDs and pfu resulted in a reduction in pfu polymerase concentration. This study provided a good start to investigate potential implications of QDs in other key molecular biology techniques. [ABSTRACT FROM PUBLISHER]

Abstract: Background: Recently, some research results showed that the circulating DNA in serum or plasma had potential for the molecular diagnosis and prognosis of certain cancers. Several methods have been employed for the quantification of circulating DNA. However, the circulating DNA levels obtained by various methods exhibited considerable differences. Additionally, these methods were labor-extensive and time-consuming, and not suitable for the quantification of circulating DNA in numerous samples due to the use of commercial DNA extraction kits for the purification of circulating DNA. We presented a new method for the quantification of circulating DNA in sera by capillary zone electrophoresis (CZE) with laser-induced fluorescence detection (LIF). Methods: In the present work, we want to make comparison between CZE-LIF assay and real time PCR for the quantification of circulating DNA levels. Linearity, intra and inter variability of two methods were evaluated. Results: The intra and inter variability of circulating DNA quantification by real-time PCR were 7.3% and 14.92%, respectively. In CZE assay the intra and inter variability were 4.19% and 6.91%, respectively. The R.S.D. values of the same coated capillary and different coated capillaries were 5.14% and 9.02%, respectively. Our data showed that the circulating DNA levels obtained by two methods had a good correlation. Moreover, we further confirmed that blood samples collection, serum preparation and other treatment procedures had a significant impact on the DNA levels in sera. Conclusion: Our data further illustrated that CZE-LIF is a simple, rapid and sensitive method for the quantification of circulating DNA in human sera, and well suitable for the analysis of a large number of samples in clinical diagnosis. [Copyright &y& Elsevier]

Activation mechanism of the nanoplatform AuNCs@MnO 2 nanosheets for GSH detection and intracellular imaging. [Display omitted] • A novel GSH-switched fluorescence nanoprobe has been constructed based on the integration between the inner filter effect of MnO 2 nanosheets and GSH-induced enhanced fluorescence of AuNCs. • The nanoplatform exhibits a high sensitivity for the detection of GSH with a low limit detection of 68 nM. • The nanoplatform achieves outstanding selectivity towards intracellular GSH over cysteine, homocysteine and ascorbic acid. • And that, it can selectively monitoring the GSH in human serum and the fluorescence imaging of endogenous GSH in living cell. Glutathione (GSH) plays a critical role in biological defense system and is associated with numerous human pathologies. However, it still remains a challenge for fluorescent detection of GSH over cysteine (Cys) and homocysteine (Hcy) because of their similar structures. In this work, MnO 2 nanosheets can efficiently quench the fluorescence of gold nanoclusters (Met-AuNCs) prepared by blending methionine and HAuCl 4 owing to their superior absorption capability. However, GSH can reduce MnO 2 nanosheets into Mn2+ which leads to the fluorescence recovery of Met-AuNCs. More intriguingly, GSH can dramatically and selectively enhance the fluorescence intensity of Met-AuNCs. Hence, a low background, ultrasensitive fluorescent detection of GSH was obtained with a detection limit of 68 nM. Moreover, the assay has been successfully used for GSH detection in human serum samples and cellular imaging with high selectivity over Cys and Hcy. [ABSTRACT FROM AUTHOR]

A fluorometric assay was constructed for supersensitive determination of Cu2+ and Co2+ based on their quenching effect on the orange fluorescence of methionine-capped gold nanoclusters (Met-AuNCs). A simple one-step method was developed for the preparation of the Met-AuNCs, employing L-methionine as both a reducing and protecting reagent. Within 10 min, water soluble Met-AuNCs were obtained with an average size of 2.4 nm. Under photoexcitation at 370 nm, the Met-AuNCs possess a maximum emission at 580 nm and a quantum yield of 2.3%. The response is fast (1 min), and the selectivity for Cu2+ and Co2+ is high over other metal ions. The detection limits for Cu2+ and Co2+ are around 47 and 420 pM, respectively. The effects were used to design a test paper for visual detection of Cu2+ and Co2+. Using this test paper, 1 μM of Cu2+/Co2+ can be detect under the UV lamp (365 nm excitation). It is perceived to be a promising tool for the rapid on-site determination of Cu2+ and Co2+ in real water samples. [ABSTRACT FROM AUTHOR]

A method is described for the determination of the activity of endonuclease. It based on the deaggregation of gold nanoparticles (AuNPs) aggregated by the action of poly(diallyldimethylammonium chloride) (PDDA). A single-stranded DNA (ssDNA) is released after enzymatic cleavage catalyzed by endonuclease. The released fragments bind electrostatically to PDDA and inhibit the PDDA-induced aggregation of AuNPs. This is accompanied by a color change from blue to red and a decrease in the absorption ratio (A630/A520). Under the optimal conditions, this ratio increases linearly in the 0.001 to 1 U·μL−1 EcoRI endonuclease activity range. The detection limit is of 2 × 10−4 U·μL−1 which is much better or at least comparable to previous reports. The method is deemed to have wide scope in that it may be used to study other endonuclease activity (such as BamHI) by simply changing the specific recognition site of the hairpin-like DNA probe. The assay may also be employed to screening for inhibitors of EcoRI endonuclease.Schematic presentation of the colorimetric assay based on the deaggregation of AuNPs for the detection of endonuclease activity. A single-stranded sequence (ssDNA) is released by the EcoRI cleavage, which electrostatically binds to PDDA and inhibits the PDDA-induced aggregation of AuNPs accompanying with a color change from blue to red. [ABSTRACT FROM AUTHOR]

A colorimetric method is described for the determination of Pt(II). It is based on the use of gold nanoparticles (AuNPs) which are known to aggregate in the presence of a cationic polymer such as poly(diallyldimethylammonium chloride) (PDDA). If, however, a mismatched aptamer (AA) electrostatically binds to PDDA, aggregation is prevented. Upon the addition of Pt(II), it will bind to the aptamer and induce the formation of a hairpin structure. Hence, interaction between aptamer and PDDA is suppressed and PDDA will induce the aggregation of the AuNPs. This is accompanied by a color change from red to blue. The effect can be observed with bare eyes and quantified by colorimetry via measurement of the ratio of absorbances at 610 nm and 520 nm. Response is linear in the 0.24-2 μM Pt(II) concentration range, and the detection limit is 58 nM. The assay is completed within 15 min and selective for Pt(II) even in the presence of other metal ions. It was successfully applied to the rapid determination of Pt(II) in spiked soil samples.Schematic representation of the method for detection of Pt(II) based on the use of a cationic polymer and gold nanoparticles. In the presence of Pt(II), aptamer interacts with the Pt(II) and prevents the interaction between aptamer and cationic polymer. Hence, cationic polymer induce the aggregation of the AuNPs and lead to the color change from red to blue. [ABSTRACT FROM AUTHOR]

Abstract: In this paper, we present a universal, highly efficient and sensitive method for the characterization of quantum dot (QD) bioconjugates based on capillary electrophoresis with laser-induced fluorescent (LIF) detection. We first prepared CdTe QDs in aqueous phase by a chemical route with mercaptopropionic acid as a ligand, and then were coupled to certain proteins using bifunctional linkage reagent or electrostatic attraction. The QD bioconjugates were characterized by capillary electrophoresis with LIF detection. We found that QD bioconjugates were efficiently separated with free QDs by the optimization of buffer pH. Furthermore, we found that ultrafiltration was an effective and simple approach to purify QD conjugates with bovine serum albumin (BSA). Due to their broad absorption spectra and size dependent emission wavelength tunability, QDs can be excited to emit different colour fluorescence using a single wavelength laser source, and therefore, we believe that CE with LIF detection will become a universal and efficient tool for the characterization of QD bioconjugates. [Copyright &y& Elsevier]

Scallop Chlamys farreri is an important aquaculture species in northern China. However, its mass mortality caused by several pathogens can result in great economic loss and negative impacts to the sustainable development of the scallop industry. Thus, improving the overall understanding of immune response mechanisms involved in host–pathogen interactions is necessary. Ferritins are conserved molecules in organisms that are involved in diverse biological processes, such as mediating host-pathogen responses. In this study, we report a novel ferritin gene from C. farreri (denoted as CfFER ). The full length of CfFER is 848 bp and contains a 5′-UTR of 113 bp, a 3′-UTR of 219 bp, and a complete open reading frame (ORF) of 516 bp. The ORF encodes a polypeptide of 171 amino acid residues with a molecular weight of approximately 19.95 kDa and an isoelectric point of 5.07. The CfFER protein exhibited typical ferritin structures, namely, a ferroxidase diiron center, a ferrihydrite nucleation center, and an iron-binding response signature. Phylogenetic analysis revealed that CfFER was closely related to other mollusk ferritin proteins. Expression of CfFER in different tissues was analyzed by quantitative real-time PCR, and results showed that CfFER was ubiquitously expressed in all examined tissues. The highest and lowest expression levels of CfFER were measured in the muscle and hemocyte, respectively. The relative mRNA expression of CfFER in response to bacterial ( Vibrio anguillarum ) and viral (acute viral necrobiotic virus) challenges sharply increased by ca. 5-fold about12 h post-infection (hpi) and then normalized at 48 hpi. Western blot analysis with polyclonal antibodies generated from the recombinant product of CfFER also demonstrated the presence of ferritin protein in hemocytes. These findings strongly suggest that CfFER is involved in the immune response of C. farreri and protection against pathogen challenge. [ABSTRACT FROM AUTHOR]