MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Jiayi Zhang,Zichun Song,Jiuying Cui,Wenbo Li,Song Huang,Bo Ling,Kai Zhang,Xianjiu Liao,Jihua Wei
The human gastrointestinal system contains the conditional pathogen Escherichia coli ( E. coli ), which is a significant cause of vomiting and diarrhea—both of which can be fatal in extreme cases. Given the surge in hospital infections and water pollution, the development of early detection and diagnosis procedures is imperative. This study proposes an electrochemical biosensor for E. coli detection, utilizing nucleic acid aptamers as recognition elements and using E. coli-2571 as an example. It is based on the CRISPR/Cas gene-editing technology. Specifically, the collateral cleavage activity of Cas12a shifts the electrochemical signal after loading the DNA sequence tagged with an electrochemical signal onto the gold electrode, which ultimately enables the identification of E. coli . The biosensor has a detection limit of 13.1 CFU/mL and exhibits a good linear relationship in the concentration range of 10 2 ––10 7 CFU/mL. It was effectively used for the quantitative detection of E. coli in milk and pure water, with a recovery rate ranging from 93.12 % to 98.93 %. The biosensor offers a rapid, precise, affordable, and sensitive tool for E. coli detection. It is anticipated to become a powerful instrument for identifying microorganisms that pose a risk to public health and human health.
研究领域:
分子生物学
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微生物学
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生物传感技术
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公共健康
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Zhina Wu,Yingtong Wang,Yaqin Zhang,Jingzheng Yi,Yutong Li,Jiasi Wang,Min Hu,Di Wang
Periodontitis is one of the most prevalent oral diseases, and the diagnosis and monitoring of periodontitis rely on clinical and imaging examinations. Matrix metalloproteinase-8 (MMP-8) in saliva is a reliable indicator of periodontitis. The traditional detection method is enzyme-linked immunosorbent assay (ELISA), which requires professional expertise and is not sensitive enough to low-abundance protein detection. In this study, we developed a nanoimmunoassay combined with clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR associated protein (Cas)12a for ultrasensitive detection of MMP-8 in saliva. Antibody-DNA-gold nanoparticles (AuNPs) and trans-cleavage activity of CRISPR/Cas12a were employed for signal amplification. With this protocol, the limit of detection (LoD) for MMP-8 was reduced to the fg/mL, indicating 100-fold higher sensitivity than that of the commercial ELISA kit and its amenability for ultrasensitive detection of trace proteins. The diagnostic sensitivity of the assay was 100 %, and the specificity was 83.3 % based on the testing of 23 clinical saliva samples. The sensitivity and specificity were considerably higher than those for ELISA. Thus, this immuno-CRISPR/Cas12a assay holds promise for both diagnosing and monitoring periodontitis in clinical settings.
研究领域:
口腔科学
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分子生物学
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生物技术
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临床诊断
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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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.
研究领域:
分子生物学
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肿瘤学
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诊断学
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产品关联
DOI
高影响因子
Advanced Science
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2025
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IF 14.1
Shuai Wu,Yincheng Liu,Tianyu Zeng,Tianci Zhou,Yanting Sun,Ying Deng,Juan Zhang,Genxi Li,Yongmei Yin
Continued development of clustered regularly interspaced short palindromic repeats (CRISPR)-powered biosensing system on the electrochemical interface is vital for accurate and timely diagnosis in clinical practice. Herein, an electrochemical biosensor based on manganese metal-organic frameworks (MOFs)-enhanced CRISPR (MME-CRISPR) is proposed that enables the efficient detection of circulating tumor DNA (ctDNA). In this design, customized enzyme stimulants (Mn 2+ ) are co-assembled with Cas12a/crRNA to form enzyme-MOF composites, which can be released quickly under mild conditions. The MOFs-induced proximity effect can continuously provide adequate Mn 2+ to sufficiently interact with Cas12a/crRNA during the release process, enhancing the trans-cleavage activity of complex available for biosensor construction. The MOFs-based enzyme biocomposites also afford efficient protection against various external stimulus. It is demonstrated that the developed biosensor can achieve ultrasensitive detection of epidermal growth factor receptor L858R mutation in ctDNA with a low detection limit of 0.28 f m without pre-amplification. Furthermore, the engineered mismatch crRNA enables the biosensor based on MME-CRISPR to detect single nucleotide variant with a high signal-to-noise ratio. More importantly, it has been successfully used to detect the targets in clinical practice, requiring low-dose samples and a short time. This strategy is believed to shed new light on the applications of cancer diagnosis, treatment, and surveillance.
研究领域:
生物医学工程
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分子诊断学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
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IF 8
Sirui Liu,Xinlan Bian,Furong Zhao,Min Cao,Zhaoyang Shang,Bingzhi Li,Xing Zhang
High mannose is overexpressed on the cell surface during carcinogenesis; however, there are currently no efficient techniques for its detection. Herein, we propose an assay called RADAR ( r emote high m a nnose d etection on cell surf a ce with C R ISPR) for the quantification of high mannose, leveraging the integration of high mannose-binding aptamer and CRISPR technique. The high mannose-binding aptamer is hybridized with a locker, which is released upon specific binding to high mannose. The released locker then activates CRISPR/Cas12a to produce a fluorescence signal. The "remote" concept in RADAR refers to the recognition of high mannose occurring on the cell surface, while signal generation takes place in a solution separate from the target. This feature makes RADAR suitable for the detection of suspended bio-samples without the need for sophisticated imaging. RADAR can detect high mannose Man 9 with a limit of detection of 2.5 nM, distinguish cancer cells from non-cancerous cells, and profile high mannose level in human serum for potential non-small cell lung cancer (NSCLC) diagnosis. This work represents the first attempt to integrate CRISPR technology into the aptasensing of high mannose, which may serve as a synthetic biology-enabled tool in glycochemistry and glycobiology.
研究领域:
合成生物学
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糖化学
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糖生物学
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生物技术
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癌症诊断
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产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2025
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IF 7.7
Kai Shi,Jiaxuan Chen,Ying Cheng,Jiuhua Song,Yang Li,Xin Cheng,Xiaofeng Bai,Jiali Chang,Tao Jiang
Microplastic (MP) pollution has attracted worldwide attention due to its harmful effects on human health and ecosystems. Herein, we develop a novel label-free electrochemical biosensor for sensitive and selective detection of MP based on split gRNA with CRISPR/Cas12a (SCas12a)-mediated cascade strand displacement and an aptamer (termed as CRISPR-MP). Within the CRISPR-MP system, the target polyvinyl chloride (PVC)/polystyrene (PS) triggers the SCas12a-mediated cascade strand displacement, and then activates the DNase activity of SCas12a. Upon cleaving the DNA probe of the G-quadruplex/hemin complex on the gold electrode, the active Cas12a triggers a decrease in the electrochemical signal.This electrochemical aptasensor shows a detection limit of 37 ng/mL of PVC and 45 ng/mL of PS, respectively. The developed CRISPR-MP system expands the utility of the CRISPR for MP detection and exhibits significant promise in protecting human health and monitoring the environment.
研究领域:
生物分析化学
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环境监测
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分子诊断学
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生物传感器技术
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纳米毒理学
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产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
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2024
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IF 4.9
Yafang Shen,Fei Jia,Yawen He,Yaping Peng,Yingchun Fu,Weihuan Fang,Yanbin Li
Rapid detection methods are urgently needed to help prevent the spread of Salmonella among food products. Herein, a CRISPR-Cas12a-powered, quantum dot-based and magnetic nanoparticle-assisted (QD-CRISPR-MNP) biosensor was proposed for the screening of Salmonella. The QD-CRISPR-MNP biosensor was designed and fabricated relying on the CRISPR-Cas12a system for better guidance of the coupling between magnetic nanoparticles (MNPs) and quantum dots (QDs). In this biosensor, the activity of the CRISPR-Cas12a system was unlocked by Salmonella and displayed collateral cleavage activity towards linker DNAs, which inhibited the coupling of the MNPs and QDs. Then, Salmonella could be quantitatively detected by simply measuring the fluorescence emitted by the QDs on the MNPs surface. This biosensor showed good detection performance for Salmonella with a detection limit of 86 CFU mL −1 , and exhibited a satisfactory specificity as well. Moreover, it overcomes the low fluorescence quantum yields and the possible high fluorescence background noise in conventional dye-based CRISPR detection platforms, acting as a powerful alternative for the detection of foodborne pathogens.
研究领域:
生物技术
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纳米技术
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食品科学
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微生物学
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产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Jiaying Wang,Yu Tian,Kai Zhang,Jiantong Jiao,Yi Zhao
NF-κB p50 is a critical transcription factor that plays an essential role in regulating immune responses, inflammatory pathways, and tumor progression. Its aberrant expression is closely linked to various pathological conditions, making it a valuable biomarker for early disease diagnosis and therapeutic monitoring. In this work, we developed a highly sensitive and reusable electrochemical biosensor capable of detecting NF-κB p50 at ultralow concentrations. The biosensor integrates a pH-responsive regeneration mechanism and a multi-stage signal amplification cascade involving Exonuclease III (Exo III)-assisted recycling, CRISPR/Cas12a-mediated trans -cleavage, and DNA tetrahedron nanostructure-functionalized electrodes. This synergistic design significantly enhances detection sensitivity and stability. The platform offers a broad dynamic detection range from 5 aM to 100 fM with a remarkably low detection limit of 2.38 aM. More importantly, the biosensor demonstrates excellent reusability through efficient regeneration under alkaline conditions (pH 10.0), with minimal loss in signal output. Validation in tenfold-diluted human serum samples yielded high recovery rates (99.61 %–106.23 %) and low relative standard deviations, confirming its practical applicability. This study not only presents a robust approach for ultrasensitive protein detection but also introduces a generalizable strategy for the regenerative electrochemical analysis of protein-based biological macromolecules in clinical diagnostics.
研究领域:
生物医学工程
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分子诊断学
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纳米技术
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产品关联
DOI
高影响因子
TALANTA
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2025
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IF 6.1
Jingwen Zhang,Xiru Zhang,Xinna Xie,Biru Han,Feng Zhao,Xinyan Yang,Wei Zhang,Yujun Jiang,Xianlong Zhang
The problem of mycotoxin contamination in foodstuffs has attracted widespread attention and posed a great threat to human health. Therefore, the sensitive and effective detection of mycotoxins is of great importance for preserving public health worldwide. In this study, a CRISPR/Cas12a-mediated marker-free fluorescent biosensing platform was constructed for highly sensitive and fast detection of aflatoxin B1. The copper nanoclusters were synthesized with marker-free DNA single strands within 5 min, showing the outstanding fluorescence properties. With the existence of aflatoxin B1, the released complementary DNA (cDNA) triggered multiple isothermal amplification reaction. Subsequently, the obtained amplification products triggered the trans -cleavage activity of CRISPR/Cas12a system, which degraded the DNA single strands for synthesis of copper nanoclusters, leading to a decreased fluorescent signal. Benefiting from good fluorescence properties of copper nanoclusters, the established biosensing platform for aflatoxin B1 detection had a high specificity and a limit of detection of 47.51 pg/mL in the linear range of 0.05–10 ng/mL. The proposed platform provided a new insight for the detection of non-nucleic acid targets.
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2024
·
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.
研究领域:
环境科学
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分子生物学
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分析化学
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
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2023
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IF 7.4
Liping Xia,Juxin Yin,Jianjian Zhuang,Weihong Yin,Zheyu Zou,Ying Mu
Rapid and sensitive pathogen detection methods are critical for disease diagnosis and treatment. RPA-CRISPR/Cas12 systems have displayed remarkable potential in pathogen detection. A self-priming digital PCR chip is a powerful and attractive tool for nucleic detection. However, the application of the RPA-CRISPR/Cas12 system to the self-priming chip still has great challenges due to the problems of protein adsorption and two-step detection mode of RPA-CRISPR/Cas12. In this study, an adsorption-free self-priming digital chip was developed and a direct digital dual-crRNAs (3D) assay was established based on the chip for ultrasensitive detection of pathogens. This 3D assay combined the advantages of rapid amplification of RPA, specific cleavage of Cas12a, accurate quantification of digital PCR, and point-of-care testing (POCT) of microfluidics, enabling accurate and reliable digital absolute quantification of Salmonella in POCT. Our method can provide a good linear relationship of Salmonella detection in the range from 2.58 × 101 to 2.58 × 104 cells/mL with a limit of detection ∼0.2 cells/mL within 30 min in a digital chip by targeting the invA gene of Salmonella. Moreover, the assay could directly detect Salmonella in milk without nucleic acid extraction. Therefore, the 3D assay has the significant potential to provide accurate and rapid pathogen detection in POCT. This study provides a powerful nucleic detection platform and facilitates the application of CRISPR/Cas-assisted detection and microfluidic chips.
研究领域:
分子生物学
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生物技术
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微生物学
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分析化学
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即时诊断
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2021
·
IF 7.46
Baoqing Zhou,Qinghua Ye,Fan Li,Xinran Xiang,Yuting Shang,Chufang Wang,Yanna Shao,Liang Xue,Jumei Zhang,Juan Wang,Yu Ding,Moutong Chen,Qingping Wu
Rapid, accurate point-of-care testing (POCT) for pathogenic bacteria detection is the key for avoiding foodborne diseases caused by pathogens or their toxins. The lateral flow biosensor (LFB) based on clustered regularly interspaced short palindromic repeats (CRISPR/Cas) has displayed remarkable potential for pathogen diagnosis. In this study, we report a novel CRISPR/Cas12a-based fluorescence enhanced LFB in conjunction with functionalized quantum dots , combined with recombinase-assisted amplification (RAA), to establish low-cost, simple, and sensitive detection of Staphylococcus aureus , namely CRA-LFB (CRISPR/Cas-recombinase-assisted amplification based LFB). The CRA-LFB assay is characterized by Cas12a-mediated trans-cleavage activation induced by the target DNA to digest biotin-DNA probes, caused no complementarity to the capture probe immobilized on the test (T) line, and resulted in an undetectable T line fluorescence signal on LFB. The naked eye or fluorescence strip reader was used to determine the fluorescence intensity of the T and control lines. The limit of detection (at optimal conditions) was as low as 75 aM of genomic DNA, and 5.4 × 10 2 cfu/mL of S. aureus in pure cultures were detected. Moreover, this CRA-LFB assay can rapidly and accurately detect S. aureus in spiked and natural meat and vegetable samples. The CRA-LFB method yielded high specificity and no interference from other nontargeted bacteria. The results exhibited high-resolution, high-intensity fluorescent signals obtained within 70 min. This bacterial detection platform is simple, low cost, almost equipment-free, and fully qualified for food and clinical diagnosis onsite testing requirements.
研究领域:
生物技术
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微生物学
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食品安全
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即时诊断
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DOI
高影响因子
危险品化学品经营许可证（带存储）