ACS Applied Materials & Interfaces
·
2021
·
IF 9.229
Qiong Hu,Yilin Luo,Xiaojing Cao,Zhuohua Chen,Yanyu Huang,Li Niu
Sensing of ultralow-abundance nucleic acids (NAs) is integral to medical diagnostics and pathogen screening. We present herein an electrochemical method for the highly selective and amplified sensing of NAs, using a peptide nucleic acid (PNA) recognition probe and a bioinspired electro-RAFT polymerization (BERP)-based amplification strategy. The presented method is based on the recognition of target NAs by end-tethered PNA probes, the labeling of thiocarbonylthio reversible addition–fragmentation chain transfer (RAFT) agents, and the BERP-assisted growth of ferrocenyl polymers. The dynamic growth of polymers is electrochemically regulated by the reduction of 1-methylnicotinamide (MNA) organic cations, the redox center of nicotinamide adenine dinucleotide (NAD+, coenzyme I). Specifically, electroreduction of the MNA cations causes the fragmentation of thiocarbonylthio RAFT agents into radical species, triggering the polymerization of ferrocenyl monomers, thereby recruiting plenty of ferrocene electroactive tags for amplified sensing. It is obvious that the BERP-based strategy is inexpensive and simple in operation. Benefiting from the high specificity of the PNA recognition probe and the amplified signal by the BERP-based strategy, this method is highly selective and the detection limit is as low as 0.58 fM (S/N = 3). Besides, it is applicable to the sensing of NAs in serum samples, thus showing great promise in the selective and amplified sensing of NAs.
研究领域:
医学诊断
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生物化学
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分析化学
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DOI
高影响因子
CHEMBIOCHEM
·
2020
·Vol. 21
·期 18
·2667-2675
·
IF 2.576
Mengjia Xu,Pan Fu,Shu Xing,Yang Zhao,Chao Zhao
Graphical Point-of-care cancer catching ? A novel PNA-DNA 2 triplex consisting of a homopurine PNA strand and two homopyrimidine DNA strands is reported for the first time. On the basis of this phenomenon, a simple, inexpensive colorimetric PNA-DNA 2 triple-helix molecular switch-based sensing platform for sensitive and selective detection of miRNAs from cancer cells is demonstrated. Peptide nucleic acids (PNAs), the synthetic DNA mimics that can bind to oligonucleotides to form duplexes, triplexes, and quadruplexes, could be advantageous as probes for nucleic acid sequences owing to their unique physicochemical and biochemical properties. We have found that a homopurine PNA strand could bind to two homopyrimidine DNA strands to form a PNA-DNA 2 triplex. Moreover, the cyanine dye DiSC 2 (5) could bind with high affinity to this triplex and cause a noticeable color change. On the basis of this phenomenon, we have designed a label-free colorimetric sensing platform for miRNAs from cancer cells by using a PNA-DNA 2 triple-helix molecular switch (THMS) and DiSC 2 (5). This sensing platform can detect miRNA-21 specifically with a detection limit of 0.18 nM, which is comparable to that of the THMS-mediated fluorescence sensing platform. Moreover, this colorimetric platform does not involve any chemical modification or enzymatic signal amplification, which boosts its applicability and availability at the point of care in resource-limited settings. The universality of this approach can be simply achieved by altering the sequences of the probe DNA for specific targets.
研究领域:
分子生物学
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医学诊断
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分析化学
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DOI
高影响因子
ACS Omega
·
2018
·
IF 0
Wanxing Xu,Shu Xing,Xiaojun Xu,Mengjia Xu,Pan Fu,Tingting Gao,Chao Zhao
With the help of peptide nucleic acid (PNA) and single-stranded (ss) DNA-specific nucleases, a simple, rapid, and label-free nanosensor is demonstrated for the sensitive and accurate detection of single-nucleotide polymorphisms (SNPs) in a homogeneous solution based on the light scattering (LS) of carbon nanotubes. PNAs are non-natural analogues of DNA with extraordinary stability toward enzymatic degradation, and DNAs in the fully matched DNA/PNA duplexes can be effectively protected from ssDNA-specific nucleases, whereas the DNAs in DNA/PNA duplexes involving a mismatch are cleaved into small fragments. This difference could be easily monitored by measuring the LS signal of single-walled carbon nanotubes (SWNTs) for the different binding affinities of SWNTs to ssPNA and PNA/DNA duplexes that can be used for the detection of SNPs. As a proof-of-principle analyte, the SNPs in the human tumor suppressor gene TP53 have been successfully genotyped by this method. The assay does not involve any chemical modification of probes, which is simple and universal. Moreover, the LS technique and the use of a sequence-specific PNA probe impart high sensitivity and selectivity. In principle, this assay can be used to detect any kind of gene mutations by simply changing the PNA sequences of the specific probe.
研究领域:
生物化学
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分子生物学
·
纳米技术
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DOI
ELECTROCHIMICA ACTA
·
2010
·
IF 3.325
Xianggang Liu,Xiangjin Qu,Hai Fan,Shiyun Ai,Ruixia Han
An electrochemical method for the detection of DNA hybridization using a novel electroactive, cationic, and water-soluble branched polyethyleneimine (BPEI)–cobalt(III)–phenanthroline(phen) polymeric indicator and single-stranded neutral peptide nucleic acid (PNA) probe on the Au electrode was developed. The indicator possesses some free amine groups, as well as cationic cobalt complexes in the polymer chain. It does not bind to neutral PNA capture probe alone. However, the indicator strongly interacts with the negatively charged backbone of the complementary oligonucleotide bound to the PNA probe through electrostatic interactions . The coordination spherical moieties also interact with the probe by embedding into the double-helix structure of PNA–DNA. These two interactions enable transduction of hybridization, producing a clear electrical signal in differential pulse voltammetry (DPV). The correlation against non-complementary DNA, three-base and one-base mismatch DNA was sharp, and the signal of indicator for the target DNA demonstrated a linear relationship within the concentration range of 5.0 × 10 −9 to 2.5 × 10 −7 M ( R = 0.9940) with a detection limit of 5.6 × 10 −10 M. These studies showed that the novel polymeric indicator and single-stranded PNA probe could be used to fabricate an electrochemical biosensor for DNA detection. This technique can provide an alternative route for expanding the range of detection methods available for DNA hybridization.
研究领域:
生物化学
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分析化学
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分子生物学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Shiwei Xu,Guofeng Wang,Yueyue Feng,Juanjuan Zheng,Liying Huang,Jiahao Liu,Yisha Jiang,Yajun Wang,Nannan Liu
MicroRNAs (miRNAs) are endogenous and noncoding single-stranded RNA molecules with a length of approximately 18–25 nucleotides, which play an undeniable role in early cancer screening. Therefore, it is very important to develop an ultrasensitive and highly specific method for detecting miRNAs. Here, we present a bottom-up assembly approach for modifying glass microtubes with silica nanowires (SiNWs) and develop a label-free sensing platform for miRNA-21 detection. The three-dimensional (3D) networks formed by SiNWs make them abundant and highly accessible sites for binding with peptide nucleic acid (PNA). As a receptor, PNA has no phosphate groups and exhibits an overall electrically neutral state, resulting in a relatively small repulsion between PNA and RNA, which can improve the hybridization efficiency. The SiNWs-filled glass microtube (SiNWs@GMT) sensor enables ultrasensitive, label-free detection of miRNA-21 with a detection limit as low as 1 aM at a detection range of 1 aM–100 nM. Noteworthy, the sensor can still detect miRNA-21 in the range of 102–108 fM in complex solutions containing 1000-fold homologous interference of miRNAs. The high anti-interference performance of the sensor enables it to specifically recognize target miRNA-21 in the presence of other miRNAs and distinguish 1-, 3-mismatch nucleotide sequences. Significantly, the sensor platform is able to detect miRNA-21 in the lysate of breast cancer cell lines (e.g., MCF-7 cells and MDA-MB-231 cells), indicating that it has good potential in the screening of early breast cancers.
研究领域:
生物医学工程
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分子生物学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 5
·1-10
·
IF 5.7
Ni Yun,Chen Xiu,Ling Chen,Zhu Ziye,Yue Yao,Wang Jie,He Aolin,Liu Ruijiang
The CYP2C19*2 gene carriers and non-carriers are closely related to the dosage of clopidogrel. To correctly guide the use of clopidogrel and promote individualized therapy, an ultra-sensitive electrochemical biosensor was developed for the detection of CYP2C19*2 gene. The heterogeneous α-Fe 2 O 3 /Fe 3 O 4 nanosheets were prepared via the hydrothermal-calcination process, and the preparation parameters were optimized. The average diameter and thickness of the nanosheets were approximately 150 nm and 53 nm, respectively; and the saturation magnetization was 80.2 emu/g. The α-Fe 2 O 3 /Fe 3 O 4 @Au nanosheets were prepared by sodium borohydride reduction method, and self-assembled to the electrode surface with magnetic field. Ultra-sensitive detection of CYP2C19*2 gene was realized through the recognition ability of strong single base mismatching of peptide nucleic acid and signal amplification effect of magnetic α-Fe 2 O 3 /Fe 3 O 4 @Au nanosheets. Under optimal detection conditions, the current had a good linear correlation with the negative logarithm of CYP2C19*2 gene concentration in the range 1 pM—1 nM, and the detection limit was 0.64 pM (S/N = 3). Meanwhile, the electrochemical signals of target DNA and incomplete complementary DNA were detected. The constructed biosensor exhibited good selectivity, reproducibility, and stability, providing a promising strategy for the detection of other gene mutations by electrochemical biosensors. Graphical Abstract
研究领域:
生物医学工程
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分子诊断学
·
材料科学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 7
·1-10
·
IF 6.232
Gao Tingting,Xing Shu,Xu Mengjia,Fu Pan,Yao Jiechen,Zhang Xiaokang,Zhao Yang,Zhao Chao
A convenient fluorometric method was developed for specific determination of DNA based on peptide nuclei acid (PNA)–regulated fluorescence resonance energy transfer (FRET) between carbon dots (CDs) and gold nanoparticles (AuNPs). In this system, CDs that display lake blue fluorescence with excitation/emission maxima at 345/445 nm were used as fluorometric reporter, while AuNPs were used as fluorescence nanoquencher. A neutral PNA probe, which is designed to recognize the target DNA, was used as a coagulant to control the dispersion and aggregation of AuNPs. Without DNA, PNA can induce immediate AuNP aggregation, thus leading to the recovery of the FRET-quenched fluorescence emission of CDs. However, the addition of the complementary target DNA can protect AuNPs from being aggregated due to the formation of DNA/PNA complexes, which subsequently produces a high fluorescence quenching efficiency of CDs by dispersed AuNPs. Under optimized conditions, quantitative evaluation of DNA was achieved in a linear range of 5–100 nM with a detection limit of 0.21 nM. This method exhibited an excellent specificity towards fully matched DNA. In addition, the application of this assay for sensitive determination of DNA in cell lysate demonstrates its potential for bioanalysis and biodetection.
A simple fluorometric biosensor for specific detection of DNA was developed based on peptide nuclei acid (PNA)–regulated fluorescence resonance energy transfer (FRET) between carbon dots (CDs) and gold nanoparticles (AuNPs).
研究领域:
分析化学
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生物化学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2019
·
IF 3.973
Shu Xing,Xiaojun Xu,Pan Fu,Mengjia Xu,Tingting Gao,Xiaokang Zhang,Chao Zhao
Rapid and sensitive single nucleotide polymorphisms (SNPs) genotyping is of particular important for early diagnosis, prevention, and treatment of specific human diseases. A simple and low-cost SNP detection method would be valuable for routine analysis in resource-limited settings. Here, we demonstrated a novel and convenient gold nanoparticle (AuNPs) based colorimetric approach for efficient screening of SNPs at room temperature without instrumentation. SNP detection is performed in a single tube with one set of unmodified AuNPs, a label-free peptide nucleic acid (PNA) probe, a single exonuclease (S1 nuclease), and the target to be tested. S1 nuclease could digest DNAs in DNA/PNA duplexes involving a mismatch into small fragments, while DNAs in the fully-matched DNA/PNA duplexes can be effectively protected by PNA from enzymatic degradation. This difference could be easily discriminated by color changes associated with gold aggregation. PNA oligomers can induce immediate AuNP aggregation even in the presence of nucleoside monophosphates (dNMPs), the digestion products of DNA . Whereas PNA/DNA duplexes can effectively stabilize unmodified AuNPs, and the stabilization effect of PNA/DNA is better than single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA). Without the need of precise temperature control and extra salt addition, SNPs are detected with a detection limit of 2.3 nM in cell lysate. Moreover, this system can effectively discriminate a range of different mismatches even in spiked cell lysate, demonstrate the potential use of this biosensor for biological samples.
研究领域:
分子生物学
·
纳米生物技术
·
医学诊断
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DOI
PubMed
高影响因子
ACS Sensors
·
2019
·
IF 6.944
Qiong Hu,Jinming Kong,Dongxue Han,Li Niu,Xueji Zhang
Sensitive and selective sensing of biological molecules is fundamental to disease diagnosis and infectious disease surveillance. Herein, an ultrasensitive and highly selective electrochemical DNA biosensor is described by exploiting the electrochemically controlled reversible addition–fragmentation chain-transfer (eRAFT) polymerization as a signal amplification strategy and the peptide nucleic acid (PNA) probes as the recognition elements. Specifically, the PNA probes with a thiol at their 5′-terminals are anchored to a gold electrode surface (via gold–sulfur self-assembly) for sequence-specific recognition of target DNA (tDNA) fragments, of which the phosphate sites serve as the anchorages for the targeted labeling (via the well-established phosphate–Zr4+–carboxylate chemistry) of the carboxyl-group-containing chain-transfer agents (CTAs) for the succedent eRAFT polymerization, wherein the initiating radicals are generated through electrochemical reduction of aryl diazonium salts under a potentiostatic condition. In the presence of ferrocenylmethyl methacrylate (FcCH═CH2) as the monomer, the grafting of polymer chains from the CTA-anchored sites as a result of the eRAFT polymerization brings numerous electroactive Fc tags to the electrode surface, outputting a high electrochemical sensing signal even in the presence of trace amounts of tDNA fragments. Under the optimized conditions, the linear range of the described electrochemical DNA biosensor spans from 10 aM to 10 pM (R2 = 0.998), with an attomolar detection limit (4.1 aM) being achieved. Moreover, the described electrochemical DNA biosensor is highly selective and applicable to the sensing of tDNA fragments in complex serum samples. Given its high efficiency, easy operation, and low cost, this biosensor shows great promise in real applications.
研究领域:
生物传感
·
分子诊断
·
电化学分析
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Mengjia Xu,Xiaomeng Wang,Juanjuan Tian,Jing Chen,Xiaojie Wei,Wei Li
Rapid, sensitive and precise detection of single-nucleotide polymorphisms (SNPs) with risk association is of great significance but still remains challenging in resource-limited clinical settings. Herein, a convenient, low-cost and label-free ratiometric fluorescent sensing strategy is developed by coupling peptide nucleic acid (PNA) clamp-improved catalyzed hairpin assembly (CHA) reaction with a novel hairpin DNA-templated dual-emissive silver nanocluster (AgNC). In the presence of targets containing single-base mismatches, the sensing platform initiates the amplification and enlarges the distance between the Ag emitter pair in AgNC, resulting in a decrease of red emission accompanied by a significant optical readout. On the contrary, the PNA clamp is capable of specifically blocking the perfectly homologous counterpart in DNA sequence and thereby impeding unwanted amplification. In this study, the SNP of oncogenic Kirsten ras ( KRAS ) gene has been accurately and effectively genotyped with a detection limit of 9.6 pM. Moreover, this nanosensor can effectively discriminate a series of SNPs with vivid color change and its universality can be simply achieved by a rational design of the relative sequences. This platform performs well for both target DNA spiked cell lysate samples and PCR amplicons from standard cell lines, implying its great practical application prospect for bioanalysis and biosensors.
研究领域:
生物化学
·
分子诊断
·
纳米医学
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产品关联
DOI
高影响因子
TALANTA
·
2021
·
IF 6.057
Mengjia Xu,Shu Xing,Yang Zhao,Chao Zhao
Here, taking the advantage of single-stranded (ss) DNA specific nuclease (S1) and peptide nucleic acid (PNA), we demonstrated a novel, rapid, and label-free colorimetric nanosensor for the sensitive and accurate detection of SNPs based on the intrinsic peroxidase-like activity of hemin-functionalized single-walled carbon nanotubes (hemin-SWCNTs). PNA, a man-made mimic of DNA with extraordinary stability toward enzymatic degradation, can effectively protect DNA in the fully matched DNA/PNA duplexes from nuclease digestion. While the DNA in DNA/PNA duplexes containing a mismatch can be cleaved into small fragments. This difference can be visually monitored from the specific color change of TMB/H 2 O 2 system by employing the peroxidase activity of hemin-SWCNTs because of its different aggregation states responding to ssPNA or DNA/PNA duplex. Under optimized conditions, the SNPs in the human tumor suppressor gene TP53 have been successfully genotyped in a linear range of 50–1000 nM with a detection limit of 0.11 nM. Moreover, this platform can effectively discriminate a series of single-base mismatches. This assay avoids the assistance of sophisticated instruments and complicated modifications of probes or nanomaterials , and function well for both cell lysate samples and PCR amplicons from standard cell lines, implying its potential practical applications for bioanalysis and biosensors.
研究领域:
生物化学
·
分子生物学
·
纳米生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Qiong Hu,Dongxue Han,Shiyu Gan,Yu Bao,Li Niu
Sensitive detection of biomolecules is integral for biomarker screening and early diagnosis. Herein, surface-initiated reversible-addition–fragmentation-chain-transfer (SI-RAFT) polymerization is exploited as a novel amplification strategy for highly sensitive electrochemical biosensing of DNA. Briefly, thiol-terminated peptide nucleic acid (PNA) probes are first self-assembled onto a gold electrode for the specific capture of target-DNA fragments; the carboxyl-group-containing dithiobenzoate 4-cyano-4-(phenylcarbonothioylthio)pentanoic acid (CPAD) is then tethered to the hybridized PNA–DNA heteroduplexes by means of the well-established carboxylate–Zr4+–phosphate chemistry and serves as the chain-transfer agent (CTAs) for subsequent SI-RAFT polymerization, which is thermally initiated in the presence of 2,2′-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (VA-044) as the free-radical initiator and ferrocenylmethyl methacrylate (FcMMA) as the monomer. Through SI-RAFT polymerization, one target-DNA fragment can be labeled by a large number of electroactive Fc tags, giving rise to significant amplification of the electrochemical signal. The SI-RAFT-polymerization-based strategy does not involve the use of natural enzymes or complex nanomaterials, offering the benefits of low cost and easy operation. Under optimal conditions, the electrochemical signal is linearly related to the logarithm of the concentration of target DNA over the range from 10 aM to 10 pM (R2 = 0.997), with a detection limit down to 3.2 aM, which is much lower than those of other amplification-by-polymerization-based methods. By virtue of its easy operation, low cost, and high efficiency, the SI-RAFT-polymerization-based amplification strategy is believed to have great application prospects in the sensitive detection of biomolecules.
研究领域:
生物化学
·
分析化学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
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