A One-Step Highly Sensitive Method for DNA Detection Using Dynamic Light Scattering

Dai, Qinggang; Liu, Xiong; Coutts, Janelle L.; Austin, Lauren A.; Huo, Qun

doi:10.1021/ja801947e

Cited by 176 publications

(146 citation statements)

References 16 publications

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“…Therefore, our method showed an ultra-wide detection range from 10 fM to 100 nM, which was wider than the values obtained by other methods. 2,5,16,26 In addition, the fluorescence intensity signal change (F -F0) increased proportionally with the concentrations of target DNA in the 1 to 100 nM range (inset in Fig. 1 based on a 3α/slope, which was better than, or comparable to, the values obtained by other methods (Table S2).…”

Section: Rapid Communicationssupporting

confidence: 48%

“…[2][3][4][5] These methods can be broadly classified into two main categories: indirect detection approaches by template replication amplification and direct detection approaches by detecting original DNA. The indirect detection approaches, such as the polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), 6 rolling-cycle amplification (RCA), 7 and nucleic acid sequence-based amplification (NASBA), 8 increase the risk of cross-contamination from amplicons and are prone to false positives arising by artifactual amplification.…”

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confidence: 99%

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A Label-free mid Turn-on Fluorescence Strategy for DNA Detection with a Wide Detection Range Based on Exonuclease III-aided Target Recycling Amplification

et al. 2017

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9The highly sensitive and selective detection of specific DNA sequences is critical for clinical diagnosis, gene therapy, mutation analysis and pathogen detection. 1 Currently, a series of analytical methods have been developed to detect DNA. [2][3][4][5] These methods can be broadly classified into two main categories: indirect detection approaches by template replication amplification and direct detection approaches by detecting original DNA. The indirect detection approaches, such as the polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), 6 rolling-cycle amplification (RCA), 7 and nucleic acid sequence-based amplification (NASBA), 8 increase the risk of cross-contamination from amplicons and are prone to false positives arising by artifactual amplification. The directdetection approaches include DNA microarray, 9 molecular beacons, 10 nicking endonuclease signal amplification (NESA), 11,12 exonuclease III (Exo III) signal amplification, [13][14][15] and so on. DNA microarrays and molecular beacons usually have lower sensitivity due to the relatively low signal amplification. NESA requires target DNA with a specific sequence for enzyme recognition that widely limits applications. Exo III does not require any specific recognition sites, thus improving its application range and flexibility. Therefore, the method of Exo III signal amplification has great potential to be developed into a more universal method for the sensitive detection of DNA.Recently, a series of analytical methods has been developed for DNA detecting based on Exo III signal amplification. However, these methods have often required labled fluorescent groups, 14 modified electrodes 13 or a turn-off mode 16 with inconvenience or high cost for applications. Furthermore, these methods often had a narrow detection range. It was inconvenient to detect the unknown concentrations of DNA. There are few simple methods for the sensitive detection of DNA with an ultra-wide detection range. Thus, a label-free, turn-on and convenient method with a wide detection range is in urgent need.The G-quadruplex structure is a tertiary structure of DNA formed by a G-rich nucleic acid sequence. 17 Interestingly, this structure can be selectively recognized by NMM, which is a commercially available unsymmetrical anionic porphyrin compound. 18,19 The fluorescence intensity of NMM is weak, but exhibits a dramatic enhancement upon binding to the G-quadruplex structure. 20,21 Recent research progress has shown that the NMM/G-quadruplex DNA system can be utilized as an excellent and label-free signal reporter to detect heavy metal ions, 22 NAD + , 23 DNA 24 and anti-cancer drugs. 25 Inspired by these reports, we report on a simple label-free and turn-on fluorescence method to detect DNA through Exo III signal amplification and the NMM/G-quadruplex DNA system.The principle of the proposed fluorescence method used to detect DNA is illustrated in Scheme 1. We have designed a stem-loop structure of a DNA probe (sequence shown in Table S1) that contains...

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Section: Rapid Communicationssupporting

confidence: 48%

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confidence: 99%

A Label-free mid Turn-on Fluorescence Strategy for DNA Detection with a Wide Detection Range Based on Exonuclease III-aided Target Recycling Amplification

et al. 2017

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“…We and many other groups have previously demonstrated that DLS can be used as a very convenient and powerful tool to monitor specific binding and non-specific adsorption of proteins to spherical gold nanoparticles [28][29][30][31]. Based on the nanoparticle size change, we and others have developed a novel platform technology, nanoparticle-enabled dynamic light scattering assay (NanoDLSay™) for biological and chemical detection and analysis with high to ultrahigh sensitivity and excellent reproducibility [32][33][34][35][36][37][38]. In this work, we applied the DLS technique to study the gold nanorods and nanorod-protein interactions.…”

Section: Introductionmentioning

confidence: 99%

Dynamic light scattering for gold nanorod size characterization and study of nanorod–protein interactions

2012

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In recent years, there has been considerable interest and research activity in using gold nanoparticle materials for biomedical applications including biomolecular detection, bioimaging, drug delivery, and photothermal therapy. In order to apply gold nanoparticles in the real biological world, we need to have a better understanding of the potential interactions between gold nanoparticle materials and biomolecules in vivo and in vitro. Here, we report the use of dynamic light scattering (DLS) for gold nanorods characterization and nanorod-protein interaction study. In the size distribution diagram, gold nanorods with certain aspect ratios exhibit two size distribution peaks, one with an average hydrodynamic diameter at 5-7 nm, and one at 70-80 nm. The small size peak is attributed to the rotational diffusion of the nanorods instead of an actual dimension of the nanorods. When proteins are adsorbed to the gold nanorods, the average particle size of the nanorods increases and the rotational diffusion-related size distribution peak also changes dramatically. We examined the interaction between four different proteins, bovine serum albumin, human serum albumin, immunoglobulin G, and immunoglobulin A (IgA) with four gold nanorods that have the same diameter but different aspect ratios. From this study, we found that protein adsorption to gold nanorods is strongly dependent on the aspect ratio of the nanorods, and varies significantly from protein to protein. The two serum albumin proteins caused nanorod aggregation upon interaction with the nanorods, while the two immunoglobulin proteins formed a stable protein corona on the nanorod surface without causing significant nanorod aggregation. This study demonstrates that DLS is a valuable tool for nanorod characterization. It reveals information complementary to molecular spectroscopic techniques on gold nanorod-protein interactions.

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“…Dynamic light scattering (DLS), a technique used routinely for determination of particle size and size distribution, has also been adopted for particle agglutination-based assays. [11][12][13] Recently, a washing-free one-step homogeneous assay protocol was implemented using gold nanoparticles by measuring the hydrodynamic diameter of the nanoparticle aggregates with a DLS instrument, and a detection limit of 0.5 ng/ml for mouse IgG was achieved. 12 However, DLS instruments are relatively delicate and sensitive to temperature, which may limit its applications in point-of-care testing and hinder its integration with microfluidic chip.…”

Section: Introductionmentioning

confidence: 99%

Homogeneous agglutination assay based on micro-chip sheathless flow cytometry

Zhang

Cheng

et al. 2015

Biomicrofluidics

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Homogeneous assays possess important advantages that no washing or physical separation is required, contributing to robust protocols and easy implementation which ensures potential point-of-care applications. Optimizing the detection strategy to reduce the number of reagents used and simplify the detection device is desirable. A method of homogeneous bead-agglutination assay based on micro-chip sheathless flow cytometry has been developed. The detection processes include mixing the capture-probe conjugated beads with an analyte containing sample, followed by flowing the reaction mixtures through the micro-chip sheathless flow cytometric device. The analyte concentrations were detected by counting the proportion of monomers in the reaction mixtures. Streptavidin-coated magnetic beads and biotinylated bovine serum albumin (bBSA) were used as a model system to verify the method, and detection limits of 0.15 pM and 1.5 pM for bBSA were achieved, using commercial Calibur and the developed micro-chip sheathless flow cytometric device, respectively. The setup of the micro-chip sheathless flow cytometric device is significantly simple; meanwhile, the system maintains relatively high sensitivity, which mainly benefits from the application of forward scattering to distinguish aggregates from monomers. The micro-chip sheathless flow cytometric device for bead agglutination detection provides us with a promising method for versatile immunoassays on microfluidic platforms.

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A One-Step Highly Sensitive Method for DNA Detection Using Dynamic Light Scattering

Cited by 176 publications

References 16 publications

A Label-free mid Turn-on Fluorescence Strategy for DNA Detection with a Wide Detection Range Based on Exonuclease III-aided Target Recycling Amplification

A Label-free mid Turn-on Fluorescence Strategy for DNA Detection with a Wide Detection Range Based on Exonuclease III-aided Target Recycling Amplification

Dynamic light scattering for gold nanorod size characterization and study of nanorod–protein interactions

Homogeneous agglutination assay based on micro-chip sheathless flow cytometry

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