Food Bioscience
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2026
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IF 6.2
Lihua Fan,Mingying Hu,Zongyuan Ou,Fan Li,Yiheng Shi,Di Wu,Yongning Wu,Guoliang Li
Sensitive, rapid, and reliable detection of aflatoxin B1 (AFB1) in food is of great importance for ensuring food safety. In this study, a dual-mode paper-based biosensing platform was developed by integrating the CRISPR/Cas12a nucleic acid cleavage system with a nanozyme-responsive DNA hydrogel for highly sensitive and portable detection of AFB1. Upon the presence of the target, AFB1 binds specifically to the aptamer, thereby activating the CRISPR/Cas12a system and subsequently inducing the degradation of the hydrogel network, which leads to the release of the encapsulated Au@Pt nanozyme. The released nanozyme catalyzes the oxidation of TMB in the presence of H 2 O 2 to generate a blue-colored product, while simultaneously producing a quantifiable diffusion signal on the paper-based substrate, enabling dual-mode colorimetric and distance-based detection. The detection limit of naked-eye detection is 0.1 ng mL −1 , and it is 0.075 ng mL −1 in the distance detection mode. The proposed method can complete the signal output within 30 min and exhibits the advantages of operational simplicity, rapid response, and high sensitivity. In practical peanut samples, satisfactory recoveries and good stability were achieved, demonstrating its potential as a promising strategy for on-site detection of trace mycotoxins in food.
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高影响因子
npj Science of Food
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2026
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IF 9.7
Wang Juan,Li Xiang,Zhang Jing,Zhao Chao,Shi Xuening
Microbial Contamination serves as an important issue in dairy safety. CRISPR/Cas12a-based methods possess tremendous potential, but are limited by nucleic acid pretreatment and specialized operations. Herein, a photothermal nanomaterial-assisted enhanced One-Tube CRISPR/Cas12a system was established to detect S. typhimurium in milk . S. typhimurium was lysed through the photothermal effect to obtain genomic DNA. A multi-crRNA strategy enables rapid signal accumulation, and the whole system was integrated into a single tube via trehalose-based aqueous biphasic systems. The proposed method requires only one step of sample addition to achieve sensitive detection within 40 min, with an LOD of 10 CFU/mL, showing potential in application.
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MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Yi Duan,Meilin Liu,Dianhui Men,Yiyao Luo,Xuhui Xia,Li Wang,Fei Deng,Danqun Huo,Changjun Hou
CA15–3 is a vital biomarker for monitoring and evaluating treatment efficacy in breast cancer patients, especially those with advanced-stage disease. Conventional immunoassays face limitations, including high cost, limited sensitivity, and dependence on potentially error-prone single-fluorescence signal outputs. To address these challenges, we designed a dual-signal-emitting sensor based on CdTe quantum dots@ZIF-8 nanomaterials, incorporating both the CRISPR/Cas12a system and catalytic hairpin assembly (CHA) technology. The binding of CA15–3 to its specific aptamer inhibits the CHA cascade reaction, thereby preventing the generation of double-stranded DNA activators necessary to initiate Cas12a-mediated trans-cleavage activity. As a result, FAM-ssDNA is extensively adsorbed onto the CdTe QDs@ZIF-8 surface, where efficient electron transfer effectively quenches its fluorescence. The incorporation of the aptamer facilitates dual-signal amplification through both the CHA and CRISPR/Cas12a systems, achieving a detection limit of 0.131 U/mL. This dual-signal output mechanism provides the system with an internal reference function, thereby significantly improving the reliability of the results. Overall, this work establishes a novel tumor biomarker detection platform that combines high sensitivity with excellent robustness.
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ACS Sensors
·
2026
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IF 10.9
Shuai Liu,Zishan Ding,Xing Lu,Zhiyong Liu,Wenrui Ma,Hanqing Xu,Hengjun Zhang,Xianling Dai,Man Shen,Yingchun Huang,Mingxuan Gao,Jing Bao,Ming Chen
Rapid and sensitive detection of antibiotic-resistant bacteria (ARB) remains a critical challenge in clinical and public health settings. This study describes the successful construction of a portable DNA activator-triggered entropy-driven catalysis-modulated CRISPR/Cas12a-based sensor (PSDA) for the ultrasensitive and rapid detection of multiple ARB. This PSDA platform utilizes a CRISPR/Cas12a-mediated signal transduction strategy, in which a target-specific DNA activator initiates an entropy-driven dynamic DNA network for signal amplification. To further enhance detection performance, a 3D-printed microfluidic chip device with a smartphone-based readout system has been integrated into the sensor, using green-emitting Zn2GeO4:Mn persistent luminescent nanoparticles as a novel molecular beacon for fluorescence enhancement. This platform enables the simultaneous detection of methicillin-resistant Staphylococcus aureus, carbapenem-resistant Pseudomonas aeruginosa, and Klebsiella pneumoniae carbapenemase 2 (KPC-2)-expressing Klebsiella pneumoniae (KPC-2 KP) with a broad dynamic range (1−107 CFU/mL), an ultralow detection limit (1 CFU/mL), and rapid analysis (∼45 min). The assay results are also highly consistent with those of conventional plate counting methods (95.48−115.15%). Overall, this study presents a cost-effective, rapid-response biosensing platform for the simultaneous detection of multiple ARB, with direct applications in clinical diagnostics, food safety monitoring, and environmental surveillance.
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高影响因子
International Journal of Nanomedicine
·
2026
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IF 8.7
Jian Tao,Binbin Zeng,Caili Bi,Minyi Xu,Xiaxian Gu,Dingding Jing,Jun Sun
Purpose Ultra-sensitive detection of exosomal miR-21 is crucial for the early diagnosis of colorectal cancer. This study aims to develop a cascade amplification platform based on surface-enhanced Raman scattering (SERS), combining rolling circle amplification (RCA) and CRISPR/Cas12a, to enable ultra-sensitive quantitative detection of exosomal miR-21.Methods A SERS substrate was constructed using streptavidin-modified concave gold nanocubes (CGNs), and gold double-stranded probes loaded with Raman reporter molecules were used as SERS probes. The two components are controllably linked via a single-stranded DNA molecule with a thiol group at one end and biotin at the other, forming a stable detection system. Target-triggered RCA generates a repeat sequence, which, under the mediation of crRNA, activates Cas12a cleavage, causing the probe to be released from the substrate surface and resulting in a significant attenuation of the SERS signal.Results This platform achieves three-stage cascading signal amplification via RCA, CRISPR/Cas12a, and SERS, with a detection limit at the femto-mole (fM) level and a total detection time of approximately 80 minutes. The results were consistent with those of qRT-PCR, demonstrating high sensitivity, high specificity, and reliability.Conclusion This study proposes an RCA-CRISPR-SERS cascade amplification platform that provides a highly sensitive and specific method for detecting miR-21 in exosomes, offering potential clinical value for the early diagnosis and dynamic monitoring of colorectal cancer.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2025
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IF 7.7
Yayun Jiang,E Jianfei,Youwei Li,Jian Chen,Jie Yang,Yang He,Gang Mai,Laibao Zheng
Group B Streptococcus (GBS) is a major cause of neonatal morbidity and mortality, underscoring the urgent need for rapid, accurate, and deployable diagnostics. Conventional culture and nucleic acid amplification tests are limited by long turnaround time, insufficient sensitivity, or dependence on sophisticated instrumentation, restricting their use in resource-limited settings. Here, we present a magnetically controlled dual-mode biosensing platform, termed c olorimetric/ p hotothermal–CRISPR/ Cas – Au @ Pt nanozyme (CP-CasAuPt), which integrates the sequence-specific trans-cleavage activity of CRISPR/Cas12a with the peroxidase-mimicking activity of Au@PtNPs. The catalytic oxidation of 3,3′,5,5′-Tetramethylbenzidine (TMB) generates oxidized TMB (oxTMB), which provides a visible colorimetric change and a near-infrared photothermal response, thereby enabling reliable dual-mode readouts with minimal instrumentation. Magnetic bead-assisted separation further reduces background signals, ensuring robust performance in complex matrices. Using GBS as a proof-of-concept, CP-CasAuPt achieved a detection limit of 10 3 CFU/mL after loop-mediated isothermal amplification, with results fully consistent with qPCR across 20 clinical samples. Beyond high sensitivity and specificity, this study establishes a generalizable strategy for multimodal molecular diagnostics, where orthogonal dual signals allow cross-validation, ensuring robust detection across diverse assay conditions. The CP-CasAuPt system thus offers a promising paradigm for rapid, instrument-free pathogen detection, particularly suited for resource-limited settings.
研究领域:
分子诊断
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生物传感器
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纳米技术
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CRISPR技术
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即时检测
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传染病检测
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2023
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IF 8.4
Cheng Qin,Xinlan Bian,Wei Lv,Bingzhi Li
CRISPR/Cas12a-based biosensors typically output fluorescent, colorimetric, and electrochemical signals, relying on reporters that suffer from low sensitivity, high cost, and low stability. Herein, we introduced a label-free chemiluminescent (CL) reporting system based on a DNAzyme known as “Supernova”. Supernova catalyzes the translocation of phosphate from a 1,2-dioxetane substrate (specifically, CDP-Star) to its 5′-OH ends, thereby triggering a CL reaction that produces light. Upon activation by the target, Cas12a cleaves Supernova, rendering it inactive in reacting with CDP-Star. Without pre-amplification, this biosensor can detect synthetic DNA corresponding to the SARS-CoV-2 N gene and monkeypox virus (MPXV) F3L gene with detection limits (LOD) of 1.5 and 2.4 pM, respectively. Integration with pre-amplification further enables the detection of SARS-CoV-2 and MPXV genes down to 1 copy/μL in real samples. Furthermore, the application of this CL system was expanded to the detection of non-nucleic acids with the integration of ATP aptamers, resulting in improved sensitivity in ATP biosensing compared to fluorescent assays. Overall, this work represents the first development of a label-free CL Cas12a reporting system, demonstrating its enhanced performance and broad applicability in synthetic biology-enabled sensing technology.
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SENSORS AND ACTUATORS B-CHEMICAL
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2022
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IF 9.221
Ailing Su,Yuan Liu,Xiumian Cao,Weiqing Xu,Chongyang Liang,Shuping Xu
A new gene detection platform based on surface-enhanced Raman spectroscopy (SERS) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system is developed, which is applied for virus gene detections. CRISPR/Cas12a activated by CRISPR ribonucleic acid (crRNA) can recognize and lock the complementary target double-stranded deoxyribonucleic acid (dsDNA), followed by the cis -cleavage on the dsDNA and the trans -cleavage on the nearby linker single-stranded DNA (ssDNA). Target-induced trans -ssDNA cleavage leads to a failure in an aggregation behavior of the designed SERS probe pair that was bridged by the linker ssDNA, which causes the SERS intensity reduced relative to the control trial that gives much stronger SERS due to the severe aggregation state of SERS probes caused by intact linker ssDNA. This sensing method achieves the highly sensitive detections of human papillomavirus (HPV) genes in serum and pseudovirus within 40 min. The detection sensitivity of this CRISPR/Cas-SERS platform can reach pM. Owing to the flexible designs of the DNA probe and crRNA, this CRISPR/Cas-SERS sensing system is a universal gene detection tool, which can be applied for the in-vitro diagnosis field and point-of-care testing.
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TALANTA
·
2022
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IF 6.556
Siwenjie Qian,Yanju Chen,Cheng Peng,Xiaofu Wang,Hui Wu,Yang Che,Huanying Wang,Junfeng Xu,Jian Wu
African swine fever virus (ASFV) can cause highly contagious and fatal disease among domestic pigs, resulting in considerable economic losses for swine breeders. There is a strong demand for accurate, rapid, and simple detection methods especially for on-site application. Nucleic acid testing is the most commonly used method for ASFVdetection. However, traditional nucleic acid purification step is time- and labor-consuming. The nucleic acid purification, amplification and amplicons detection rely on laboratory settings which limits the on-site detection. Here, we proposed a simple and cost-effective detection method that utilized filter paper to purify nucleic acids from swine blood and employed CRISPR/Cas12a-mediated loop-mediated isothermal amplification (LAMP) reaction to detect ASFV. The filter paper which was made into dipsticks could effectively purify nucleic acids from whole blood in 2 min. This simple and low-cost purification method avoided multiple pipetting steps and potential amplification inhibitors (e.g., ethanol) that were generally used in traditional nucleic acids extraction processes. After nucleic acid purification, the lyophilized LAMP reagent dissolved by elution solution was employed to perform isothermal amplification reaction on a portable heating block. The CRISPR/Cas12a system was designed to specifically detect amplicons. Assisted by a portable homemade device, the fluorescent signals produced by positive samples could be observed by the naked eye, while negative samples remained colorless. The whole detection procedure could be finished within 50 min with a detection limit of one copies/μL. This established method provided a novel strategy for rapid visualized detection and showed great potential for on-site application.
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高影响因子
ACS Synthetic Biology
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2022
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IF 5.249
Chunfeng Feng,Wenbin Liang,Fei Liu,Yu Xiong,Man Chen,Pan Feng,Mingjing Guo,Yunxia Wang,Zhongjun Li,Liqun Zhang
The mutation status of epidermal growth factor receptor (EGFR) exon 19 is of great importance for predicting sensitivity to tyrosine kinase inhibitors (TKIs) in the treatment of non-small-cell lung cancer (NSCLC). However, the development of simple, sensitive, and no-nonspecific amplification platforms for EGFR 19del detection in NSCLC remains a challenge. Herein, we developed a novel, simple, and highly sensitive naked-eye assay utilizing CRISPR/Cas12a-triggered no-nonspecific nucleic acid amplification (NAA) with rolling circle amplification (RCA) as a model for EGFR 19del detection. Typically, circular padlocks are designed to be the trans-cleavage substrate of Cas12a/crRNA and serve as templates for RCA. Since the target EGFR 19del induces robust trans-cleavage activity of the Cas12a/crRNA duplex, the surrounding circular padlocks are cleaved into random short linear fragments that are unable to initiate RCA, resulting in a colorless solution. However, in the absence of EGFR 19del, the inactivated Cas12a enzymes cannot cleave the circular padlocks, and they remain able to serve as templates to initiate RCA to generate long single-stranded DNA to further fold into G-quadruplex/hemin DNAzymes to catalyze the oxidation of 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS2–), generating a color response that is obvious to the naked eye. As expected, this strategy with a detection limit as low as 20 fM exhibited robust selectivity and anti-interference ability. Moreover, this method was applicable for detecting EGFR 19del in real serum samples and showed high consistency with real-time quantitative polymerase chain reaction (qPCR) and sequencing results, providing a promising strategy for the early noninvasive diagnosis and guidance of clinical treatment for cancer.
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JOURNAL OF HAZARDOUS MATERIALS
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2024
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IF 12.2
Manyan Qiu,Zhiyu Yuan,Nan Li,Xinyan Yang,Xianlong Zhang,Yujun Jiang,Qianyu Zhao,Chaoxin Man
Sensitive detection and point-of-care test of bacterial pathogens is of great significance in safeguarding the public health worldwide. Inspired by the characteristics of horseradish peroxidase (HRP), we synthesized a hybrid nanoflower with peroxidase-like activity via a three-component self-assembled strategy. Interestingly, the prepared nanozyme not only could act as an alternative to HRP for colorimetric biosensing, but also function as a unique signal probe that could be recognized by a pregnancy test strip. By combining the bifunctional properties of hybrid nanoflower, isothermal amplification of LAMP, and the specific recognition and non-specific cleavage properties of CRISPR/Cas12a system, the dual-readout CRISPR/Cas12a biosensor was developed for sensitive and rapid detection of Salmonella enterica . Moreover, this platform in the detection of Salmonella enterica had limits of detection of 1 cfu/mL (colorimetric assay) in the linear range of 10 1 –10 8 cfu/mL and 10 2 cfu/mL (lateral flow assay) in the linear range of 10 2 –10 8 cfu/mL, respectively. Furthermore, the developed biosensor exhibited good recoveries in the spiked samples (lake water and milk) with varying concentrations of Salmonella enterica . This work provides new insights for the design of multifunctional nanozyme and the development of innovative dual-readout CRISPR/Cas system-based biosensing platform for the detection of pathogens. Environmental implication The presence of bacterial pathogens is pervasive in the environment and poses a significant threat to public health safety. To prevent foodborne disease outbreaks, it is crucial to develop sensitive and reliable pathogen detection strategies for rapid and on-site detection of pathogens in both environmental and food samples. Herein, we developed a novel dual-readout CRISPR/Cas biosensing platform based on nanozyme with bifunctional properties and loop-mediated isothermal amplification (LAMP) for highly sensitive and on-site detection of Salmonella enterica in lake water and milk samples. The proposed method has good stability and high accuracy in the detection of Salmonella enterica .
研究领域:
生物技术
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纳米技术
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病原体检测
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生物传感器
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即时诊断
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高影响因子
ANALYTICAL CHEMISTRY
·
2025
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IF 6.7
Chen Huang,Ling-Jie Guo,Jia-Yi Li,Yue-Wen Chen,Bing Wu,Yi-Wei Liu,Chen-Zhi Zhang,Wei-Wei Lin,Ye Yang,Jin-Yuan Chen,Zhou-Jie Liu
The combination of CRISPR-Cas12a with aptamers can potentially improve the sensitivity, specificity, and speed of non-nucleic acid target detection. Nevertheless, current CRISPR-Cas12a aptasensors, solely dependent on aptamer affinity and overlooking the enzymatic regulation of CRISPR-Cas12a, may produce false-positive signals. We proposed a CRISPR-Cas12a aptasensor synergistically regulated by an aptamer and RNA, which incorporated aptamer-mediated drug recognition and CRISPR-powered signal amplification in a one-pot format. Herein, a regulatory RNA probe synergized with the conformational switching of a drug-induced aptamer, enabling precise regulation of Cas12a trans-cleavage activity via toehold-mediated strand displacement (TMSD). This dual regulatory mechanism transformed the aptamer-activated CRISPR-Cas12a sensing process into a TMSD-driven conditional reaction, avoiding false-positive signals and thus achieving better detection performance with a lower detection limit. With the vancomycin (VCM) aptamer as a model, the aptasensor can detect VCM within 30 min from 2% serum, 1% synovial fluid, and 1% cerebrospinal fluid, with a detection limit of 13.62, 7.56, and 6.75 nM, respectively. The proposed aptasensor was incorporated into a custom 3D-printed portable fluorometer and underwent clinical validation in 22 VCM serum samples, reporting no significant difference when compared with the enzyme-multiplied immunoassay technique, confirming the reliability for point-of-care quantification. It further received cross-validation with a quinine aptamer, suggesting universality [a linear range of 10–250 nM (R2 = 0.985) and a detection limit of 0.42 nM]. By integration of aptamer selectivity with CRISPR programmability, this work presents a novel robust biosensing paradigm for point-of-care drug concentration detection.
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高影响因子
危险品化学品经营许可证（带存储）