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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ANALYTICA CHIMICA ACTA
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2026
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IF 6.1
Xinying Wang,Yujie Chang,Yiyan Zhang,Xuan Yu,Yixuan Zou,Zheqi Liu,Fuchuan Qi,Jingjing Ming
Accurate and rapid determination of aflatoxin M1 (AFM1) in food is crucial for ensuring the safety of the human diet. In view of this, we proposed a proximity-inducible Cas13a activity and hydrogel-assisted paper-based distance sensor for the detection of aflatoxin M1. The single-stranded DNA (ssDNA) and AFM1 aptamer were split into two parts and linked to the terminus of two separate DNA activator strands, respectively. By introducing AFM1, it stimulated these two DNA activator strands to assemble with CRISPR RNA (crRNA) through spatial proximity mechanism between the target and these two DNA activators to activate the Cas13a trans-cleavage towards the RNA-containing site within hairpin probe to release a DNA initiator. The released DNA initiator triggered the catalytic hairpin assembly (CHA) between magnetic beads (MBs)-DNA capture probe and trypsin-DNA detection probe. After hydrolysis of gelatin hydrogel by trypsin, water released from the hydrogel flowed along the pH indicator strip and exhibited a distinct mark. Depending on this portable method, the limit of detection (LOD) was achieved at 23.6 ng L -1 . Furthermore, our developed strategy can be applied to measure AFM1 in ultra-high temperature (UHT) milk with satisfactory effects, and its excellent specificity and reproducibility also demonstrate that our platform has great potential for application in environmental monitoring.
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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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Advanced Science
·
2026
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IF 14.1
Xuhui Fan,Ruru Zhang,Linjun Yang,Shixiong Chen,Meijuan He,Yongqiang Wang,Linjie Huang,Jianfeng Zeng,Shuwang Wu,Mingyuan Gao,Han Wang
Radiotherapy is a fundamental treatment for prostate cancer; however, its therapeutic efficacy is frequently limited by radioresistance mediated through DNA repair mechanisms and other biological factors. Although gene therapy holds promise for overcoming such resistance, identifying effective radiosensitization targets and developing efficient gene delivery systems remain practically challenging. In this study, transcriptomic analysis of radiotherapy-treated prostate cancer cells revealed a marked upregulation of DNA polymerase delta subunit 4 (POLD4), a target that has not been systematically studied. To evaluate the potential of POLD4 for overcoming radioresistance, CRISPR-Cas9-based plasmids along with ultrasmall superparamagnetic iron oxide nanoparticles (USPIONs) were encapsulated within cationic liposomes for achieving an MRI-trackable gene delivery platform (plasmid and iron oxide co-loaded liposomes, termed PIO@Lipo). Comprehensive in vitro and in vivo studies demonstrated that PIO@Lipo enabled efficient POLD4 knockdown. Furthermore, PIO@Lipo synergized with radiotherapy to induce extensive DNA damage, promote tumor cell apoptosis, and remodel the immunosuppressive microenvironment. Notably, PIO@Lipo displayed superior MRI contrast enhancement capability and passive tumor targeting ability. In conclusion, this study has identified POLD4 as a potent target for radiosensitization, capable of disrupting DNA damage-repair homeostasis through MRI-monitored gene editing, thereby offering a promising strategy to enhance the efficacy of radiotherapy in prostate cancer.
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International Journal of Nanomedicine
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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
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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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ANALYTICAL CHEMISTRY
·
2025
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IF 6.7
Muhammad Sohail,Shuyao Ma,Barira Mushtaq,Muhammad Usman Haider,Bingzhi Li,Xing Zhang,He Huang
Conventional CRISPR/Cas sensing platforms exhibit poor efficiency concerning reporter-based demerits, including their interference-labile nature, photobleaching, low robustness due to a single output signal, and probe-concentration dependence. Herein, a carbon dots (CDs)-based dual-emissive fluorescent ratiometric CRISPR/Cas reporter platform was fabricated for biosensing and other analytical applications to bottleneck the demerits of conventional reporters, integrating the benefits of a ratiometric strategy and four-element doped carbon dots (4D CDs) as a transducer. Briefly, doping enhances the optical and physicochemical traits of CDs and minimizes the effect of the interfering species. A series of state-of-the-art N, P, S, and Cu codoped CDs (4D CDs) were synthesized using the hydrothermal approach and statistical tools, such as Box-Behnken design, analysis of variance, and others, enhancing photophysical traits, surface features, and sensitivity of CDs. The red-emissive CDs were prepared by using the same procedure but different precursors. The optimum 4D CDs (blue-emissive) and red-emissive CDs were used to unleash the principle of the fluorescent ratiometric CRISPR/Cas reporter system for diverse applications. Finally, the designed 4D CDs-based CRISPR/Cas biosensor was applied for nucleic acid monitoring, such as the COVID-19 nucleic acid. This project disclosed the controlled-doping principle to synthesize 4D CDs and unleashed the mechanism of ratiometric dual-emissive CRISPR/Cas-powered reporters for precise sensing applications. We anticipate the implementation of this technology in commercial analytical, biosensing, point-of-care, and other applications.
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BIOSENSORS & BIOELECTRONICS
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2022
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IF 12.545
Yang Sun,Cheng Yang,Xiaolin Jiang,Pengbo Zhang,Shuo Chen,Fengxia Su,Hui Wang,Weiliang Liu,Xiaofei He,Lei Chen,Baoyuan Man,Zhengping Li
False detection of SARS-CoV-2 is detrimental to epidemic prevention and control. The scalar nature of the detected signal and the imperfect target recognition property of developed methods are the root causes of generating false signals. Here, we reported a collaborative system of CRISPR-Cas13a coupling with the stabilized graphene field-effect transistor, providing high-intensity vector signals for detecting SARS-CoV-2. In this collaborative system, SARS-CoV-2 RNA generates a “big subtraction” signal with a right-shifted feature, whereas any untargets cause the left-shifted characteristic signal. Thus, the false detection of SARS-CoV-2 is eliminated. High sensitivity with 0.15 copies/μL was obtained. In addition, the wide concerned instability of the graphene field-effect transistor for biosensing in solution environment was solved by the hydrophobic treatment to its substrate, which should be a milestone in advancing it's engineering application. This collaborative system characterized by the high-intensity vector signal and amazing stability significantly advances the accurate SARS-CoV-2 detection from the aspect of signal nature.
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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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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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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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