MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yuanxun Gong,Jiayi Zhang,Huiying Liang,Yanling Wang,Zichun Song,Jihua Wei,Qianli Tang,Kai Zhang,Xianjiu Liao
Matrix metalloproteinases (MMPs) are critical for the degradation of extracellular matrix components, which is essential for processes such as tissue remodeling, wound healing, and angiogenesis. MMP-2, in particular, degrades type IV collagen and is associated with diseases such as cancer and cardiovascular conditions, making it a significant biomarker. This study presents a novel electrochemical biosensor for the detection of MMP-2. The biosensor combines the specificity of a programmable RNA detection system with the amplification capabilities of T7 RNA polymerase and CRISPR/Cas12a. In the absence of MMP-2, Peptide Nucleic Acid (PNA) adsorbs on the DNA template, preventing RNA amplification and resulting in a low electrochemical signal. When MMP-2 is present, it cleaves a specific peptide, releasing the DNA template and enabling RNA synthesis. This activation leads to the cleavage of single-stranded DNA probes, increasing the electrochemical signal. The biosensor demonstrates high sensitivity with a detection limit of 0.496 fM for MMP-2 and excellent specificity, effectively distinguishing MMP-2 from other proteins. It successfully detected MMP-2 in biological samples, such as liver cancer cell supernatants, indicating its potential for clinical diagnostics. This innovative biosensor offers a powerful tool for the early detection and monitoring of diseases associated with MMP-2 activity.
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高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Yuexin Wang,Bo Shen,Nini Luo,Cai Li,Haiping Wu,Yanshuang Wang,Shen Tian,Xuemiao Li,Rui Liu,Xinmin Li,Junman Chen,Wei Cheng,Shijia Ding,Rui Chen,Meifang Xiao,Qianfeng Xia
The simple, rapid, and accurate detection of highly lethal melioidosis is crucial for early clinical diagnosis and improving cure rates. Currently, due to time consumption, low sensitivity and detection rate the existing clinical detection methods cannot satisfy the needs of clinical diagnosis. Herein, a novel self-enhanced porous hydrogel material (Au@PEI-ABEI@Pt) for ultrasensitive ECL strategy of detection B. pseudomallei was report. The novel porous hydrogel material composed of PEI-ABEI porous hydrogel, gold nanoparticles (AuNP) and platinum nanoparticles (PtNP) have large specific surface area and porous structure, which not only fix more ABEI to realize self-enhanced ECL signal amplification, but also facilitate ion diffusion and efficient utilization of catalytic materials, realizing rapid electron transfer and zero-distance catalysis, significantly improving the initial signal of ECL sensor. Moreover, the most attractive aspect is that Au@PEI-ABEI@Pt hydrogels with good biocompatibility can achieve widespread application of CRISPR/Cas12a in solid-phase carriers without affecting the sensitivity, specificity and shearing activity of CRISPR/Cas12a. After coupling with the ECL system and CRISPR/Cas12a signal amplification strategy, the Au@PEI-ABEI@Pt can achieve an ultrasensitive ECL assay of B. pseudomallei with the LOD of 5 CFU mL −1 in complex samples, with high specificity and stability to effectively classify B. pseudomallei and other Gram-negative bacteria. This study shows that the developed porous hydrogel materials not only serve as an excellent ECL signal reporter to significantly improve the detection sensitivity of ECL biosensors, but also provide a new approach for the wide application of CRISPR/Cas systems in solid-phase carriers.
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Small
·
2025
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IF 12.1
Chenggong Zhu,Qunfang Huang,Ruijie Fu,Zhen Xun,Qishui Ou,Yunlei Xianyu,Can Liu
Hepatitis B virus (HBV) infection remains a significant global public health issue, and rapid detection of HBV DNA is crucial for disease prevention and control. However, traditional methods for HBV DNA detection are limited by their reliance on precise instruments and single readout, which can hardly meet the requirements of on-site detection. In this study, the Mg 2+ -enhanced trans-cleavage activity of clustered regularly interspaced short palindromic repeats/associated protein 12a (CRISPR/Cas12a) is reported and coupled with loop-mediated isothermal amplification (LAMP) and Au@Pt nanoparticles as a signaling reporter for on-site detection of HBV DNA. This triple-modal biosensing strategy enables multiple signal readouts including UV–vis spectrum, RGB value, and temperature with high sensitivity and accuracy. The linear detection ranges using UV–vis spectroscopy, RGB color recognition, and photothermal modes are from 50 to 10,000 copies µL −1 , with the limitation of detection of 24.07, 39.65, and 23.33 copies µL −1 , respectively. This biosensing strategy is further employed for the qualitative detection of HBV DNA in 48 serum samples, achieving sensitivities of 100%, 100%, and 95.24% for triple modes. This work offers a promising tool of next-generation LAMP-CRISPR/Cas12a for the rapid and portable detection of nucleic acids.
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PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
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2025
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IF 4.6
Jiaxuan Chen,Kai Shi
Accurate detection of glyphosate (GLY) residues is essential for protecting public health, ensuring food safety, and preventing environmental pollution caused by excessive or improper herbicide application. Based on the asymmetric CRISPR/Cas12a system as a signal amplification technique, with phosphorothioate-modified hairpin G-quadruplex (psHG4) acting as a signal probe, a platform named ACC-GLY is developed for the sensitive and accurate detection of GLY. In the designed ACC-GLY platform, the target GLY specifically binds to the aptamer, and the cascading signal amplification strategy, driven by the DNase activity of a single Cas12a, is initiated by two competitive guide RNAs. Under the influence of Cas12a's DNase activity, the psHG4 probe is cleaved, releasing the psG4 sequence. The released psG4 sequence then binds to Thioflavin T (ThT), forming a complex that generates a fluorescence signal. Under optimal conditions, the detection platform can specifically detect GLY at a concentration as low as 0.3 pM. The detection platform demonstrates significant capability in detecting GLY in tap water and corn samples, highlighting its vast potential for applications in environmental monitoring and food safety. To the best of our knowledge, this is the first study to utilize the CRISPR/Cas12a system for GLY detection.
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MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Jun Wang,Xueting Cui,Jing Zhang,Juan Li,Bo Pang,Jinhua Li
Microcystin-LR (MC-LR) is a significant environmental pollutant that presents a substantial risk to public health. The monitoring with simple approaches is essential for the prevention and management of MC-LR. In this study, we present an aptasensor platform utilizing recombinase polymerase amplification (RPA)-CRISPR/Cas12a technology to accurately identify MC-LR. The MC-LR aptamers were attached to magnetic beads (MBs) following combination with cDNA by hybridization, resulting in the formation of the MB aptasensor. Subsequently, glucose oxidase (GOx)-modified MB were synthesized using single-stranded DNA (MB-ssDNA-GOx) in order to induce alterations in color upon exposure to the TMB-glucose system. The colorimetric signal was visible to the naked eye and exhibited a positive correlation with MC-LR concentration. The sensing platform described in this study demonstrated great sensitivity and achieved a limit of detection of 0.1 μg/L. Furthermore, this methodology exhibited exceptional selectivity and favorable recovery rates, hence showcasing its strong suitability for the analysis of actual water samples. Hence, the assay exhibites significant potential for future environmental monitoring.
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BIOSENSORS & BIOELECTRONICS
·
2026
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IF 11.8
Yuanqian Xiong,Li Li,Jun Huang,Xuan-ang Shen,Jiayi Sun,Yuqian Guo,Daofeng Liu,Tongtong Ma,Xiaolin Huang,Yonghua Xiong
A target DNA preamplification-free CRISPR/Cas12a sensing platform was established via TEA system. • TEA system achieves a 10 6 -fold signal enhancement compared with conventional CRISPR/Cas12a system. • TEA-FQLFA system achieves attomolar-level detection of target dsDNA (0.081 aM). • TEA-FQLFA system enables sensitive detection of Salmonella typhimurium with a LOD of 6.94 CFU/mL. The use of the CRISPR/Cas12a system combined with lateral flow assays shows significant promise for point-of-care testing in resource-limited regions. However, the use of isothermal amplification technologies to preenrich target DNA fragments inherently increases the risk of aerosol contamination during strip testing. Here, we introduce a target-triggered exponential amplification-enhanced CRISPR/Cas12a system combined with a fluorescence-quenching lateral flow assay (TEA-FQLFA) for ultrasensitive detection of Salmonella typhimurium . The TEA system converts a single Cas12a recognition event into sequential dual Cas12a activation, initiating a cleavage cascade amplification that achieves a 10 6 -fold signal enhancement without the need for a target DNA preamplification operation. Moreover, hyperbranched gold-coated polymer microspheres (HGPMs), characterized by ultrahigh extinction coefficients and a broad absorption band in the 300–800 nm range, are utilized as fluorescence quenching reporters of the FQLFA for simultaneous colorimetric visualization and fluorescence-based quantitative detection of target DNA concentration. By leveraging the synergistic integration of TEA-mediated signal amplification and HGPM-enabled optical signal enhancement, the TEA-FQLFA system achieves attomolar-level detection of target DNA (0.081 aM) and enables sensitive detection of Salmonella typhimurium in complex food matrices, with detection limits of 7.95 CFU/mL in colorimetric mode and 6.94 CFU/mL in fluorescence mode. In summary, this work establishes a versatile platform that integrates cascade signal amplification with dual-mode lateral-flow for ultrasensitive food safety detection.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2025
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IF 7.7
Zhi-Li Yao,Qing-Nan Li,Yong-Ji Li,Hao-Rui Li,Si-Wei Yang,Chen-Yu Wang,Jia-Yi Shi,Shun-Li Wu,Shuang-Qing Liu,Gui-Mei Han,Jing Wang,Li-Yong Xing,Yan Zhao,De-Ming Kong,Qi-Liang Cai
Clear cell renal cell carcinoma (ccRCC) is the most prevalent form of kidney cancer in adults, with rising global incidence and mortality rates. Early diagnosis is critical for improving patient outcomes. However, current diagnostic techniques often rely on expensive and complex instrumentation, which limits their accessibility. This creates an urgent need for affordable, sensitive, and user-friendly diagnostic methods. MiR-210–3p, a biomarker significantly upregulated in ccRCC, shows promise for non-invasive diagnostic applications. In this study, we innovatively developed a fluorescence-colorimetric dual-mode platform for miR-210–3p detection by utilizing a recently identified hemin aptamer with a stem-loop structure and by combining the trans -cleavage activity of the CRISPR/Cas12a system with the peroxidase-like activity of the aptamer/hemin complex. Moreover, using fluorescently labeled aptamer or its complementary sequence as cleavage substrates for CRISPR/Cas12a allows us to achieve two types of dual-mode detection: fluorescence “turn on” combined with colorimetric “turn off”, and fluorescence “turn on” combined with colorimetric “turn on”. This dual-mode detection platform combines sensitive quantification (with a detection limit for miR-210-3p of 0.95 pM) with rapid and visually interpretable on-site analysis, significantly enhancing the reliability and accuracy of test results. The platform has good scalability, as it is capable of detecting miR-210-3p and can also be extended to detect other tumor biomarkers. This versatility holds great potential for integration with big data analytics to personalize miRNA profiles, aiding in tumor staging, monitoring, and the development of treatment strategies, thereby advancing the field of precision medicine.
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CHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 12.5
Yongzhen Dong,Aoting Gu,Chunling Li,Huihui Wang,Yang Zhou,Zhixiong Pi,Chengjia Jiang,Yiping Chen
A CbAgo-CRISPR/Cas12a-powered cascade amplification sensing strategy was proposed. • It combined CbAgo's precise cleavage with the Cas12a's robust trans-cleavage activity. • An amplified fluorescent biosensor was developed for detecting Salmonella Typhimurium . • The proposed biosensor demonstrated comparable analytical performance to qPCR assay. Sensitive and accurate detection of foodborne pathogens constitutes a critical imperative for safeguarding food safety. Herein, we proposed a Clostridium butyricum argonaute (Cb A go)-CRISPR/ C as12a-powered synergistic amplification strategy and developed a DNA amplification-free f luorescent ( ACF ) biosensor for ultrasensitive detection of Salmonella Typhimurium . In this biosensor, magnetic beads-phage conjugates enabled the selective capture of viable Salmonella Typhimurium , followed by rapid DNA extraction within 5 min using commercial lysis buffers. A synergistic signal amplification strategy was designed for efficient biosensing by utilizing the primary cleavage product (tDNA) of the CbAgo system as a molecular bridge to integrate CbAgo and Cas12a systems. Specifically, CbAgo performed primary cleavage of target DNA to generate tDNA fragments. These tDNA subsequently activated the CRISPR/Cas12a system, initiating its trans-cleavage activity to degrade fluorescent reporter probes for quantitative analysis. This tDNA-mediated integration synergizes CbAgo's precision cleavage capability with Cas12a's efficient trans-cleavage activity, enabling exponential amplification. Under optimized conditions, the ACF biosensor exhibited a linear detection range of 10 2 –10 7 CFU/mL and a detection limit of 24.8 CFU/mL for Salmonella Typhimurium without DNA amplification steps. Moreover, the ACF biosensor demonstrated comparable analytical performance to the qPCR assay and achieved satisfactory results in the detection of Salmonella Typhimurium in 20 food samples, highlighting its potential as an ultrasensitive tool for foodborne pathogen monitoring. Download: Download high-res image (107KB) Download: Download full-size image
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BIOSENSORS & BIOELECTRONICS
·
2023
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IF 12.6
Hua Wang,Yang Sun,Yuan Zhou,Yujie Liu,Shuo Chen,Wenbo Sun,Zidong Zhang,Junqing Guo,Cheng Yang,Zhengping Li,Lei Chen
At present, the 100% case fatality and the cross-infection of virus strains make the ASFV 's harm to society continue to expand. The absence of an effective commercial vaccine poses early detection remains the most effective means of curbing ASFV infection. Here, we report a cascaded detection platform based on the CRISPR-Cas12a system combined with graphene field-effect transistor sensors. The cascade platform could detect ASFV as low as 0.5 aM within 30 min and achieve typing of wild and vaccine strains of ASFV in a single detection system. The evaluation of 16 clinical samples proved that, compared with the gold standard Real-time PCR method, this platform has outstanding advantages in sensitivity, specificity and typing. Combining CRISPR-Cas12a's high specificity with the bipolar electric field effect of graphene field-effect transistor, the cascade platform is expected to achieve clinical application in the field of DNA disease detection, and provides a new direction for multi-strain disease typing.
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SCIENCE OF THE TOTAL ENVIRONMENT
·
2023
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IF 9.8
Fang Deng,Jiafeng Pan,Manjia Chen,Zhi Liu,Junhua Chen,Chengshuai Liu
Polychlorinated biphenyls (PCBs) are ubiquitous persistent organic pollutants that cause harmful effects on environmental safety and human health. There is an urgent need to develop an intelligent method for PCBs sensing. In this work, we proposed a logic gate biosensing platform for simultaneous detection of multiple PCBs. 2,3′,5,5′-tetrachlorobiphenyl (PCB72) and 3,3′,4,4′-tetrachlorobiphenyl (PCB77) were used as the two inputs to construct biocomputing logic gates. We used 0 and 1 to encode the inputs and outputs. The aptamer was used to recognize the inputs and release the trigger DNA. A catalytic hairpin assembly (CHA) module is designed to convert and amplify each trigger DNA into multiple programmable DNA duplexes, which initiate the trans -cleavage activity of CRISPR/Cas12a for the signal output. The activated Cas12 cleaves the BHQ-Cy5 modified single-stranded DNA (ssDNA) to yield the fluorescence reporting signals. In the YES logic gate, PCB72 was used as the only input to carry out the logic operation. In the OR, AND, and INHIBIT logic gates, PCB72 and PCB77 were used as the two inputs. The output signals can be visualized by the naked eye under UV light transilluminators or quantified by a microplate reader. Our constructed biosensing platform possesses the merits of multiple combinations of inputs, intuitive digital output, and high flexibility and scalability, which holds great promise for the intelligent detection of different PCBs.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Xiaotian Guan,Jingru Zhao,Zhou Sha,Yujie Liang,Jianghong Huang,Jun Zhang,Shuqing Sun
Exosomes, as novel biomarker for liquid biopsy, exhibit huge important potential value for cancer diagnosis. However, various proteins show different expression levels on exosomal membrane, and the absolute concentration of exosomes in clinical samples is easily influenced by a number of factors. Here, we developed a CRISPR/Cas12a and aptamer-chemiluminescence based analysis (CACBA) for the relative abundance determination of tumor-related protein positive exosomes in plasma for breast cancer diagnosis. The total concentration of exosomes was determined through captured CD63 using a CRISPR/Cas12a-based method with the LoD of 8.97 × 10 3 particles/μl. Meanwhile, EpCAM and MUC1 positive exosomes were quantitatively detected by aptamer-chemiluminescence (ACL) based method with the LoD of 1.45 × 10 2 and 3.73 × 10 2 particles/μl, respectively. It showed that the percentages of EpCAM and MUC1 positive exosomes offered an excellent capability to differentiate breast cancer patients and healthy donors. The high sensitivity, strong specificity, outstanding anti-interference capability, and steady recovery rate of this approach offered higher accuracy and robustness than the commercialized method in clinical trial. In addition with good stability, easy preparation and low cost, this method not only provides a new approach to rapid analysis of exosome proteins, it may be quickly extended to the diagnoses of various cancers.
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JOURNAL OF HAZARDOUS MATERIALS
·
2025
·
IF 11.3
Haoran Zheng,Jiayu Guo,Han Wang,Yurou Wang,Wenxiang Zhou,Ke Liu,Qiang Ma,Yanfei Qi
Bacteria are prevalent environmental pollutants. Live bacteria can proliferate and spread under appropriate conditions, presenting higher risks compared to non-viable counterparts. However, detecting live bacteria remains a challenge. Adenosine triphosphate (ATP), an energy currency in organisms, offers a reliable biomarker for the live bacteria sensing. Herein, we developed a zeolitic imidazolate framework-90 (ZIF-90) enhanced ATP sensing platform to detect live bacteria. The nanosystem which based on ZIF-90, encapsulating the DNA decorated magnetic beads (MB@S1) through self-assembly. When the S. aureus aptamers on ZIF-90 bonded to bacteria, the skeleton structure of ZIF-90 was disrupted by ATP leakage from live bacteria, leading to the release of MB@S1. Then, the MB@S1 initiated the hybridization chain reaction (HCR) and they were transduced by CRISPR-Cas12a to amplify the signal twice. The proposed ZIF-90 blocking sensing strategy (ZIF-90 strategy) exhibited remarkable sensitivity with the limit of detection (LOD) down to 0.223 pM for ATP detection, about 500 folds lower than the traditional ATP aptamer blocking sensing strategy (aptamer strategy). Furthermore, we used a smartphone for on-site analysis, realizing the quantification of live S. aureus with the LOD of 2.0 CFU/mL. Therefore, the approach possessed great application potential for public health, environment monitoring, bioanalysis and food safety.
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