Enzyme-free amplified DNA assay: five orders of linearity provided by metal stable isotope detection

Liu, Yu; Ding, Yonggang; Gao, Ying; Li, Rui; Hu, Xiaorong; Lv, Yi

doi:10.1039/c8cc07036a

Cited by 21 publications

(2 citation statements)

References 32 publications

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“…Compared with spectroscopic-technique-based biosensors, mass spectrometry has the advantage of excellent specificity. However, its detection limit for direct detection of miRNA is generally in the nanomolar level. , One of the main reasons for the relatively low sensitivity of mass spectrometry for miRNAs is the low ionization efficiency of miRNAs. , Various methods including organic mass tags, metal element labeling, and isothermal amplification are used to increase the ionization efficiency and enhance the sensitivity for miRNA detection. − Among them, metal nanoparticles are quite promising mass tags offering high detection sensitivity because of the presence of many thousands of metal atoms in a single metal nanoparticle of a 10 nm diameter. − Although the sensitivity problem of mass spectrometry for miRNA detection is effectively solved by mass tagging techniques, its specificity is lost since the mass spectral signals come from the mass tags instead of miRNAs themselves, and the discrimination between the target miRNAs from nontarget ones with SNPs is based on the differences in their mass spectral intensities. Mass spectrometric methods possessing both high sensitivity and specificity are highly desired for the detection of miRNAs.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive and Specific Mass Spectrometric Platform for miRNA Detection Based on the Multiple-Metal-Nanoparticle Tagging Strategy

et al. 2021

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The multiple-metal-nanoparticle tagging strategy has generally been applied to the multiplexed detection of multiple analytes of interest such as microRNAs (miRNAs). Herein, it was used for the first time to improve both the specificity and sensitivity of a novel mass spectroscopic platform for miRNA detection. The mass spectroscopic platform was developed through the integration of the ligation reaction, hybridization chain reaction amplification, multiple-metal-nanoparticle tagging, and inductively coupled plasma mass spectrometry. The high specificity resulted from the adoption of the ligation reaction is further enhanced by the multiple-metal-nanoparticle tagging strategy. The combination of hybridization chain reaction amplification and metal nanoparticle tagging endows the proposed platform with the feature of high sensitivity. The proposed mass spectrometric platform achieved quite satisfactory quantitative results for Let-7a in real-world cell line samples with accuracy comparable to that of the real-time quantitative reverse-transcriptase polymerase chain reaction method. Its limit of detection and limit of quantification for Let-7a were experimentally determined to be about 0.5 and 10 fM, respectively. Furthermore, due to the unique way of utilizing the multiple-metal-nanoparticle tagging strategy, the proposed platform can unambiguously discriminate between the target miRNA and nontarget ones with single-nucleotide polymorphisms based on their response patterns defined by the relative mass spectral intensities among the multiple tagged metal elements and can also provide location information of the mismatched bases. Its unique advantages over conventional miRNA detection methods make the proposed platform a promising and alternative tool in the fields of clinical diagnosis and biomedical research.

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Section: Introductionmentioning

confidence: 99%

Highly Sensitive and Specific Mass Spectrometric Platform for miRNA Detection Based on the Multiple-Metal-Nanoparticle Tagging Strategy

et al. 2021

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“…As a method of choice for longtime stable analysis, ICPMS is prevalent in metal elements and metal nanoparticles research, thanks to its longtime signal stability, high sensitivity, wide dynamic range, and absolute quantification feature traceable to primary SI units . ICPMS has been widely used for the measurement of metal containing nanoparticles, metal isotopes, metal elements, and biomolecules after metal stable isotope tagging. − Besides excellent luminescent properties, the DNA-templeted CuNPs intrinsically comprising a large amount of copper isotopes ( 63 Cu and 65 Cu), which can be efficiently and long-term stably detected by elemental mass spectrometry . Exonuclease I (Exo I), a widely used 3′-termini specific exonulease for ssDNA, was selected as a modal analyte.…”

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confidence: 99%

Label-Free Nuclease Assay with Long-Term Stability

Liu

Chen

et al. 2019

Anal. Chem.

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Nature endows us with a unique toolbox of highly specific enzymes, while their detection is of great importance in biological processes. The label-free assay based on DNA-templated CuNPs is widely accepted for enzyme assay owning to its simple procedure, fast kinetic, high quantum yield, and large Stokes shift. A challenge in the application of them is the low fluorescent signal stability of DNA-templated CuNPs, whose signal sharply decreases in a few minutes after formation. In this work, a long-term stable nuclease assay is proposed by utilizing the elemental mass spectrometry detection of CuNPs. The high sensitivity was also realized, thanks to a great number of copper isotopes (63Cu and 65Cu) intrinsically incorporated in CuNPs. The experimental conditions, including the length of polyT ssDNA template, the concentration of polyT template, the concentration of Cu2+, the sodium ascorbate concentration, the copper reduction reaction time, and the Exonuclease I (Exo I) digestion reaction time, were investigated in detail. The dynamic range of the Exo I concentration from 0.1 U/mL to 20 U/μL was obtained using inductive coupled plasma mass spectrometry (ICPMS) 63Cu signal, with a detection limit (3σ) of 0.029 U/mL. The ICPMS 63Cu signal remained unchanged for at least 18 days. The spiked-recovery assay in RPMI 1640 cell medium also demonstrated the reliability of the proposed nuclease assay.

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Ratiometric DNA Walking Machine for Accurate and Amplified Bioassay

Wang

Liu

et al. 2019

Chemistry A European J

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DNA walkingm achines openedn ew avenues for the biosensing and demonstrated great successi nthe past few years.S inceD NA machines are mainly nonequilibrium systemsd riven by dynamic interactions, the matrix effects on DNA machines is ab ottlenecka nd more intricate than common DNA-mediated assays,e speciallyf or complicated physiological samples. Herein, to realizea n accurate and reliable quantitative machine, ar atiometric DNA walking machinew as developedi nh uman serums and cell lysatesb ased on the elemental isotoper atio measurement. The target DNA-triggered walking machine converted and amplified biological signals into mass spectrometric signal ratios ( 197 Au/ 115 In) via ab urnt-bridge mechanism. Under the optimized conditions, the limit of detection (LOD, 3s)w as 8f Mf or target DNA,w ith ad ynamic linear range of 0.05-0.7 pM. The ratiometric DNA walking machine was directly appliedi nh uman serum samples with satisfactory recoveries of 94 to 105 %, demonstrating an excellent stability and ah igh accuracy.C ombining the aptamer-based specific recognition,t he proposedD NA machine is expected to be av ersatile platform for other targets,s uch as small biomolecules and proteins.Supporting information and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

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Enzyme-free amplified DNA assay: five orders of linearity provided by metal stable isotope detection

Abstract: The metal stable isotope detection strategy demonstrates a wide linear range and a low detection limit, showing promising potential in clinical diagnosis.

Cited by 21 publications

References 32 publications

Highly Sensitive and Specific Mass Spectrometric Platform for miRNA Detection Based on the Multiple-Metal-Nanoparticle Tagging Strategy

Highly Sensitive and Specific Mass Spectrometric Platform for miRNA Detection Based on the Multiple-Metal-Nanoparticle Tagging Strategy

Label-Free Nuclease Assay with Long-Term Stability

Ratiometric DNA Walking Machine for Accurate and Amplified Bioassay

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