SENSORS AND ACTUATORS B-CHEMICAL
·
2023
·
IF 8.4
Pei Wang,Bohui Zhou,Ziye Liu,Yining Zhao,Robert Chunhua Zhao,Jinhui Peng,Juan Zhang
The accurate detection of immune checkpoint molecules on the surface of tumor cells is of great significance for immunotherapy. Herein, we propose a method based on cellular interface supported cascade signal amplification mediated by CRISPR/Cas13a system for in situ analysis of immune checkpoint molecules on bladder cancer cells. CD47 and EpCAM on bladder cancer cells can be sensitively monitored through T7 transcription amplification, CRISPR/Cas13a system and HCR triple signal amplification. Moreover, the established method can specifically differentiate EpCAM and CD47 positive bladder cancer cells and well serve for the evaluation of two kinds of membrane protein inhibitors. In a word, the method we proposed can successfully realize in situ analysis of immune checkpoint molecules on bladder cancer cells for guiding immunotherapy.
研究领域:
癌症生物学
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分子生物学
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免疫学
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生物技术
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产品关联
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Longfei Tang,Jingyi Yang,Ziting Liu,Qin Mi,Lili Niu,Juan Zhang
A new nanoprobe (DTAAT) has been well designed based on DNA tetrahedron supported hybridization chain reaction (HCR) and further applied for in situ analysis of bacteria. Antibody-aptamer conjugate (AA) has been coupled with DNA tetrahedron (DT) followed by the linkage of trigger strand (A). Bacteria can be preciously recognized by dual binding of both antibody and aptamer in DTAAT, to release trigger strand which is complementary with aptamer. By virtue of proximity effect, the released trigger strand can further efficiently induce the simultaneous occurrence of arrays of HCR at three vertices of DT, with rapid reaction rate of 359.29 min −1 in comparison with that of 212.60 min −1 for free HCR. DTAAT can be served for in situ imaging of bacteria by confocal laser scanning microscopy and single-cell characterization by flow cytometry. Moreover, using DTAAT as probe, the quantitative analysis of bacteria can be realized with high specificity and anti-interference capability. In a word, the developed DTAAT nanoprobe has a great potential for accurate and sensitive in situ analysis of bacteria.
研究领域:
分子生物学
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分析化学
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生物医学工程
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微生物学
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Yifeng Lan,Yingqi Ma,Yingwen Xu,Yuan Zhang,Rujia Deng,Lulu Liu,Boya Hou,Haiyan Cui,Keming Yun,Zhiwen Wei,Yanli Wei
Effective truncation and mutation can obtain highly active nucleic acid aptamers for the target, especially avoiding non-specific binding, uncertainty to the tertiary structure and low binding affinity of the lengthy aptamer. In this work, a new aptamer, T-AF28 (28 nt) with better affinity to AFB1 was obtained by truncating 22 bases and mutating 3 bases of the reported AFB1 aptamer (50 nt), and the thermodynamic information, structural changes and binding domains of aptamer T-AF28 in the presence of AFB1 or AFM1 were further investigated by isothermal titration calorimetry, circular dichroism, and molecular dynamics. On this basis, Using optimized aptamers as recognition elements, a novel label-free and enzyme-free fluorescent aptasensor for AFB1 and AFM1 was developed by synergistic amplification effect of hybridization chain reaction (HCR) coupled molybdenum disulfide nanosheets and copper nanoparticles in situ synthesized (CuNPs) on an extra-long double-stranded template generated by HCR as the fluorescent probe. Under the optimal experimental conditions, sensitive and selective fluorescent detection of AFB1 and AFM1 was achieved with wide linear detection ranges of 0.030 ng/mL-46.8 ng/mL and 0.030 ng/mL-32.8 ng/mL, respectively. And this aptasensor can be applied to accurate detection of AFB1 and AFM1 in grains, edible oil, tea, and milk with the recovery in the range of 85.20 %-117.0 %. This work not only provides a theoretical basis for the design of AFB1 and AFM1 aptasensors and expands the application of AFB1 aptasensors in the field of food safety, but also breaks a new path for the development of HCR-MoS 2 Ns CuNPs aptasensors.
研究领域:
食品安全
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分析化学
·
分子生物学
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产品关联
DOI
高影响因子
TALANTA
·
2018
·
IF 4.244
Jing Bao,Changjun Hou,Yanan Zhao,Xintong Geng,Mickey Samalo,Huisi Yang,Minghong Bian,Danqun Huo
In present study, a sensitive and effective electrochemical microRNA (miRNA) sensing platform is successfully developed by integrating gold nanoparticles/polypyrrole-reduced graphene oxide (Au/PPy-rGO), catalyzed hairpin assembly (CHA) and hybridization chain reaction (HCR) multiple signal amplification strategy. Firstly, Au/PPy–rGO was employed onto a bare GCE by electrodeposition that can greatly enhanced conductivity and effectively immobilize probes. Then, the thiolated capture probes (SH-CP) were self-assembled on the Au/PPy–rGO modified GCE via Au-S bond. The target miRNA triggered the dynamic assembly of the two hairpin substrates (H1 and H2), leading to the cyclicality of the target miRNA and the formation of H1–H2 complexes without the assistance of enzyme. Subsequently, the newly emerging DNA fragment of H2 triggered the HCR when a mixture solution (hairpins H3 and H4) and produced dsDNA polymers. Finally, a substantial amount of methylene blue (MB) as signal indicator was intercalated into the minor groove of the long dsDNA polymers to achieve detected electrochemical signal. The fabricated sensor is able to detect miRNA-16 (model target) with concentration range from 10 fM to 5 nM with a low detection limit (LOD) of 1.57 fM (S/N = 3). Current research suggests that the developed multiple signal amplification platform has a great potential for the applications in the field of biomedical research and clinical analysis.
研究领域:
生物医学工程
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分析化学
·
纳米技术
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分子生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Zipeng Yin,Ling Zhu,Zijian Lv,Meijin Li,Dianping Tang
Persistent luminescent nanoparticles (PLNPs) are a class of materials with excellent optical properties , which can continue to emit light for a long time after removing the excitation light source. This feature enables PLNPs to be used for development of biological detection modes without autofluorescence background. In this study, we prepared Zn 2 GeO 4 : Mn 2+ , Pr 3+ (ZGOMP) nanorods through a one-pot hydrothermal method . Using the pH-responsive luminescence behavior of ZGOMP, we developed an autofluorescence-free biosensor using ZGOMP as a probe and gluconic acid as a quencher to detect prostate-specific antigen (PSA). Hybridization chain reaction (HCR) and magnetic separation system were introduced in the design to achieve efficient signal amplification. Under the optimal conditions, the as-designed autofluorescence-free sensing platform showed high selectivity , and showed a good luminescence response to PSA within the linear range of 0.001–10 ng/mL at a detection limit of 0.64 pg/mL. The excellent analytical performance shows that the current strategy provides an effective platform for clinical sample analysis.
研究领域:
生物传感
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纳米技术
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分析化学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Min-Xi Li,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu
Herein, a label free and sensitive miRNA detection method with enhanced practical applicability was developed based on the locked nucleic acid (LNA) assisted repeated fishing amplification strategy. The working mechanism of the proposed method is as follows: 1) a DNA probe (i.e, L-DNA) with LNA bases is immobilized onto the surface of a gold foil. The L-DNA hybridizes with the 3′ terminus of the first strands of complementary deoxyribonucleic acid (cDNA) of the target miRNA in the test samples; 2) The protruding 5′ terminus of the cDNA serves as a ‘fishhook’ to repeatedly fish the products of a hybridization chain reaction (HCR) out from a ‘reaction tube’; 3) The HCR products can be unloaded from the gold foil into a ‘product tube’ through temperature-controlled dehybridization; 4) The concentration of the target miRNA is determined based on the fluorescence intensity generated by the addition of SYBR-Green I (SG) into the ‘product tube’. The proposed platform was applied to the detection of miRNA-122 in cell lysate samples and obtained quantitative results with accuracy comparable to the quantitative reverse transcription PCR method (qRT-PCR). It is worth pointing out that the proposed platform achieved a limit of detection value of 2.9 fM for miRNA-122 by a simple but effective LNA-assisted repeated fishing amplification strategy instead of complicated enzyme-based amplification techniques. It is reasonable to expect that the proposed method provides a competitive alternative for designing practically applicable, cost-effective and label-free miRNA detection methods.
研究领域:
生物化学
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分子生物学
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纳米技术
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·Vol. 148
·期 11
·2459-2464
·
IF 4.2
Qianying Wang,Junyi Zhang,Ruo Yuan,Yun Xiang
The convenient and sensitive assay of antibiotic residuals is important for monitoring food safety and human health. Here, with a new self-constrained metal ion-dependent DNAzyme and hybridization chain reaction (HCR) signal amplifications, a simple label-free and highly sensitive aptamer-based fluorescent sensing assay for sarafloxacin is established. The target sarafloxacin molecules bind the aptamers in the duplex DNA probes to release the complementary strands, which cyclically activate the catalytic activity of the self-constrained DNAzymes to cut the substrate sequences to liberate many ssDNA strands. Subsequent HCR of two hairpins is initiated by these ssDNA strands for yielding long dsDNAs with lots of G-quadruplexes, which intercalate thioflavin T dye to generate drastically magnified fluorescence for non-label and sensitive sarafloxacin detection at the detection limit of 29 pM. Also, a highly selective assay of low levels of sarafloxacin in diluted milk samples has been demonstrated, showing the great potential of our approach for the development of versatile sensitive and convenient aptasensors for monitoring different antibiotics.
研究领域:
分析化学
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生物化学
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食品科学
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生物技术
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产品关联
DOI
PubMed
高影响因子
Biomolecules
·
2025
·Vol. 15
·期 9
·1275
·
IF 4.8
Yarong Zhang,Hongyan Ma,Zhenyao Gao,Miao Li,Fan Yang,Lingbo Sun,Yuecheng Zhang
Apurinic/apyrimidinic endonuclease 1 (APE1) selectively cleaves the apurinic/apyrimidinic site (AP site) in DNA, playing a critical role in base excision repair and genomic stability maintenance. Aberrant APE1 expression has been linked to numerous diseases, including cardiovascular disorders, neurological conditions, and various cancers. However, existing methods for detecting trace levels of APE1 remain suboptimal for certain applications. To address this limitation, we developed an innovative biosensing platform for ultrasensitive APE1 detection by integrating APE1-specific site recognition with hybridization chain reaction (HCR)-based signal amplification, enabling enzyme- and label-free bioassays. In this system, APE1 recognizes and cleaves the AP site-containing hairpin (HP) probe, releasing a single-stranded HCR initiator that triggers cascaded HCR amplification. Owing to the high efficiency of HCR, this method achieves ultrahigh sensitivity, with a calculated detection limit of 1.0 × 10−8U/mL. Furthermore, the biosensor demonstrates robust performance in cell lysates and is applicable for screening and evaluating APE1 inhibitors.
研究领域:
分子诊断
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生物分析化学
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癌症研究
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DNA修复机制
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生物传感器开发
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Dongxia Li,Yanan Peng,Liangqing Lu,Delun Chen,Bingrong Wang,Qiumei Pu,Qionglin Zhou,Xinxin Xiao,Mingpeng Fu,Xiaohong Wang,Jinchun Tu,Hua Pei,Qiang Wu
Adenosine triphosphate (ATP), as a crucial signaling molecule, is of high potential diagnosis value for multiple diseases. However, current technologies for ATP detection struggle to satisfy the harsh conditions required for biospecimen tests, existing several limitations for clinical practical application. In this study, the DNA tile response system was designed and constructed by integrating nucleic acid aptamer, hybrid chain reaction (HCR), and DNA tile self-assembly technology to achieve accurate detection of ATP in body fluid specimens. The DNA tile response system, possessing high-order structure characteristic, could remove HCR false-positive products and conquer the high noise background to enhance the detection sensitivity. Under the optimal reaction parameters, this system achieved the low limit of detection (LOD) of 189.23 nM over a wide linear range and splendid detection specificity faced with multiple interference terms. Furthermore, this system, possessing simple storage conditions, long-term detection stability, and superior anti-interference performance, was sufficient to satisfy the detection requirements for complicated biological specimens in multi-scenarios. Ultimately, the DNA tile response system showed excellent diagnostic efficiency that could easily distinguish urinary tract infection patients and healthy controls through ATP quantification, as well as hypertension patients and healthy ones. This system provides a high-potential rapid detection tool for early diagnosis and dynamic monitoring of urinary tract infections and hypertension, holding enormous potential for clinical translation due to the superiorities of simple operation, general applicability, and low instrument requirements.
研究领域:
生物技术
·
医学诊断
·
分子生物学
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产品关联
DOI
高影响因子
Scientific Reports
·
2020
·Vol. 10
·期 1
·1-8
·
IF 3.998
Tan Xiao-Hui,Li Yu-Bin,Liao Yan,Liu Hua-Zhong
In this work, an enzyme-free fluorescence resonance energy transfer (FRET) strategy was established for rapid and specific detection of the DNA sequence from Vibrio parahaemolyticus (VP) using hybridization chain reaction (HCR) amplification and triplex DNA. The triplex forming oligonucleotide (TFO) was labelled with carboxyfluorescein (FAM) as fluorescence donor, and hairpin sequence H1 was labelled by tetramethylrhodamine (TAMRA) as fluorescence receptor. In the present target VP DNA, the hairpin structure of molecular beacon (MB) was opened, the free end was released and hybridized with H1-TAMRA, and the HCR reaction was triggered by the alternate supplementation of H1-TAMRA and H2 to produce the notch double helix analogue. After the addition of TFO-FAM, a triplex structure was formed between HCR products (H1-TAMRA/H2) and TFO-FAM. A close contact between the donor and the receptor resulted in FRET. Under the optimal conditions, the fluorescence quenching value was inversely proportional to the concentration of target VP DNA in the range of 0.1–50 nmol L −1 , and the detection limit was 35 pmol L −1 .
研究领域:
生物化学
·
分子生物学
·
分析化学
·
微生物学
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产品关联
DOI
PubMed
高影响因子
FOOD CONTROL
·
2025
·
IF 6.3
Xinru Ren,Weiqing Sun,Bowen Li,Yijing Xiao,Xudong Yue,Meili Yang,Qing-ao Pang,Rui Zhu,Zhiqiang Guo,Haiping Zhang,Yu Wang,Su Liu,Jiadong Huang
The detection of pathogenic bacteria plays a crucial role in ensuring food safety and public health. The integration of isothermal amplification technology with the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) system offers a versatile approach to detecting foodborne pathogens. However, this method is often constrained by the incompatibility of the reaction systems and the lack of control over CRISPR/Cas activity. Here, we demonstrated a fluorescence biosensing strategy based on linear causal-regulated and autocatalysis-driven DNA circuits (termed LC@ADC) for highly sensitive and specific detection of Salmonella Typhimurium ( S. Typhimurium ). The approach of combining hybridization chain reaction (HCR) with DNAzyme-driven CRISPR/Cas12a activation was proposed for the first time, enabling not only one-step cascade signal amplification but also controllable DNA circuits. The LC@ADC strategy is capable of quantifying concentrations of S. Typhimurium ranging from 10 to 1 × 10 5 CFU/mL, with a detection limit as low as 6.53 CFU/mL. Furthermore, the LC@ADC strategy has been successfully employed for the detection of S. Typhimurium in samples of orange juice, chicken, and lake water. Therefore, this strategy provides a novel approach for the high-performance detection of pathogenic bacteria.
研究领域:
分子生物学
·
生物技术
·
食品安全
·
公共卫生
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生物传感器
·
微生物学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2024
·
IF 5.7
Zaiyong Zhang,Jinglong Li,Chunlin Chen,Yuwei Tong,Dehui Liu,Cuizhi Li,Huan Lu,Li Huang,Wanling Feng,Xiaoting Sun
Brain natriuretic peptide (BNP) is considered to be an important biomarker of heart failure (HF) attracting attention. However, its low concentration and short half-life in blood lead to a low-sensitivity detection of BNP, which is a challenge that has to be overcome. In this work, we propose a highly specific, highly sensitive T7 RNA polymerase-assisted clustered regularly interspaced short palindromic repeats (CRISPR)/Cas13a system to detect BNP via an electrochemiluminescence (ECL) sensing platform and incorporate Exo III-hairpin and dumbbell-shaped hybridization chain reaction (HCR) technologies. In this detection scheme, the ECL sensing platform possesses low background signal and high sensitivity. Firstly, the T7 promoter-initiated T7 RNA polymerase acts as a signal amplification technique to generate large amounts of RNAs that can activate CRISPR/Cas13a activity. Secondly, CRISPR/Cas13a is able to trans -cleave the surrounding trigger strand to produce DNA1. Thirdly, DNA1 is involved in the co-amplification reaction of Exo III and hairpin DNA, which subsequently triggers a dumbbell-shaped HCR technology. Eventually, a large number of Ru (II) molecules are inserted into the interstitial space of the dumbbell-shaped HCR to generate a strong ECL signal. The CRISPR/Cas13a possesses outstanding specificity for a single base and increased sensitivity. The tightly conformed dumbbell-shaped HCR provides higher sensitivity than the traditional linear HCR amplification technique. Ultimately, the clever combination of several amplification reactions enables the limit of detection (LOD) as low as 3.2 fg/mL. It showed promise for clinical sample testing, with recovery rates ranging from 98.4% to 103% in 5% human serum samples. This detection method offered a valuable tool for early HF detection, emphasizing the synergy of amplification strategies and specificity conferred by CRISPR/Cas13a technology.
研究领域:
生物技术
·
分子生物学
·
分析化学
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）