Sensors and Actuators Reports
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2025
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IF 7.6
Yanli Sun,Hongying Li,Tao Wang,Kai Zhang,Xueliang Wang
MicroRNA-17 (miR-17) is a critical biomarker linked to various cancers and diseases, underscoring the need for sensitive and selective detection methods. Herein, an ultra-sensitive electrochemiluminescent (ECL) sensor for miR-17 detection utilizing MOF-derived porous CuFe 2 O 4 nanospheres and CRISPR/Cas13a trans-cleavage-triggered hybridization chain reaction (HCR) amplification was presented. The porous CuFe 2 O 4 nanospheres possessed a high surface area and excellent catalytic activity, significantly enhancing ECL emission. To the best of our knowledge, this is the first report demonstrating that MOF-derived CuFe 2 O 4 can effectively enhance the electrochemiluminescence of Ru(bpy) 3 2 ⁺ for miR-17 detection. The CRISPR/Cas13a system provided miR-17 recognition and trans-cleavage activity, triggering HCR for signal amplification. Under optimized conditions, the biosensor exhibited a wide linear detection range from 20 aM to 5000 aM, with a detection limit of 9.23 aM. The system discriminated miR-17 from closely related miRNAs with negligible cross-reactivity, which demonstrated its outstanding selectivity. Practical applicability was confirmed by successful quantification of miR-17 in diluted human serum which achieved recovery rates between 95.2% and 104.8%. The results validated the biosensor’s potential for early disease diagnosis and clinical monitoring of miR-17-associated disorders.
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2023
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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.
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SENSORS AND ACTUATORS B-CHEMICAL
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2022
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IF 9.221
Li Shuofeng,Wang Fangfang,Chun Wang,Zhi Wang,Qiuhua Wu
As vital biomarkers for diagnosis of diseases, the anomalously expressed microRNAs (miRNAs) are closely linked to the occurrence and development of many diseases. Quantitative analysis of miRNAs has remarkable significance to clinical diagnosis and therapy of different cancers. In this study, the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) 13a (CRISPR/Cas13a) system was first exploited to directly and specifically recognize the miRNA at the guidance of CRISPR RNA (crRNA), which can activate the nonspecific trans-ribonuclease activity of Cas13a. Then, combining the CRISPR-Cas13a system with terminal deoxynucleotidyl transferase (TdT)-triggered template-free DNA extension to form the fluorescent copper nanoparticles (CuNCs), a simple but sensitive method (named as Cas13a-TFCN) was developed for highly specific detection of miRNA. Based on the CRISPR/Cas13a system coupled with TdT-triggered simple but efficient signal amplification strategy, the developed Cas13a-TFCN method showed high sensitivity for miRNA detection with the low detection limit of 0.7 fM. Furthermore, the proposed Cas13a-TFCN approach was successfully applied for accurate quantification of miRNA in complex biological samples, indicating its practical application ability. Moreover, featuring high sensitivity and simplicity without complicated probe design and fluorophore labeling, the proposed Cas13a-TFCN has a promising applications prospect in clinical diagnosis.
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BIOSENSORS & BIOELECTRONICS
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2025
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IF 10.5
Yingjing Fan,Weilun Pan,Qi Xiu,Yue Qiao,Peng Zhang,Tong Liao,Junjie Feng,Chengtao Nie,Shuai Chu,Ningcen Li,Xiuhua Wu,Lei Zheng,Zhen Cai,Bo Li
Tumor immunotherapy has significantly improved cancer treatment, yet immune checkpoint inhibitors only benefit a subset of patients. A critical challenge is non-invasive detection of tumor PD-L1 mRNA, a key biomarker that dynamically reflects PD-L1 expression and correlates with therapeutic outcomes. Current methods are hindered by the instability of circulating mRNA and the need for invasive tissue biopsies. Herein, we introduce the Cationic LIposome capsulated CRISPR-Cas13a Kit (CLICK) for in-situ detection of circulating PD-L1 mRNA in extracellular vesicles (EVs). CLICK achieves direct target identification in 10 μL plasma within 2 hours without EV purification or RNA extraction, demonstrating a detection limit of 10 3 EVs/mL for PD-L1 mRNA. By integrating membrane fusion-mediated CRISPR delivery with nanoflow cytometry, CLICK resolves PD-L1 mRNA (+) EV subpopulations at single-vesicle resolution, revealing heterogeneous distributions. Clinically validated in 53 immunotherapy patients, CLICK accurately stratifies progressive disease (PD) from stable disease (SD) patients (AUC = 0.8236; 95% CI: 0.7054–0.9417; p= 0.0006) based on PD-L1 mRNA (+) EV percentages. This work establishes a rapid, blood-based paradigm for therapy response monitoring, advancing liquid biopsy through precise EV RNA characterization.
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PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2023
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IF 13.6
Jihua Wei,Zichun Song,Jiuying Cui,Yuanxun Gong,Qianli Tang,Kai Zhang,Xinlei Song,Xianjiu Liao
In this study, we introduce an electrochemiluminescence (ECL) sensing platform based on the "Entropy-driven triggered T7 amplification-CRISPR/Cas13a system" (EDT-Cas). This platform combines a programmable entropy-driven cycling strategy, T7 RNA polymerase, and the CRISPR/Cas13a system to amplify the determination of the SARS-CoV-2 RdRp gene. The Ti 3 C 2 T x -compliant ECL signaling molecule offers unique benefits when used with the ECL sensing platform to increase the assay sensitivity and the electrode surface modifiability. To obtain the T7 promoter, the SARS-CoV-2 RdRp gene may first initiate an entropy-driven cyclic amplification response. Then, after recognizing the T7 promoter sequence on the newly created dsDNA, T7 RNA polymerase starts transcription, resulting in the production of many single-stranded RNAs (ssRNAs), which in turn trigger the action of CRISPR/Cas13a. Finally, Cas13a/crRNA identifies the transcribed ssRNA. When it cleaves the ssRNA, many DNA reporter probes carrying -U-U- are cleaved on the electrode surface, increasing the ECL signal and allowing for the rapid and highly sensitive detection of SARS-CoV-2. With a detection limit of 7.39 aM, our method enables us to locate the SARS-CoV-2 RdRp gene in clinical samples. The detection method also demonstrates excellent repeatability and stability. The SARS-CoV-2 RdRp gene was discovered using the "Entropy-driven triggered T7 amplification-CRISPR/Cas13a system" (EDT-Cas). The developed ECL test had excellent recoveries in pharyngeal swabs and environmental samples. It is anticipated to offer an early clinical diagnosis of SARS-CoV-2 and further control the spread of the pandemic.
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PubMed
高影响因子
TALANTA
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2025
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IF 6.1
Haixiang Li,Shuo Lian,Zhiwei Zhang,Weiye Bi,Qingyou Meng,Qingwei Ding
Brain natriuretic peptide (BNP) is considered a reliable marker of heart failure disease, and its timely detection can provide important pathological information to prevent or treat heart failure. In this article, an electrochemiluminescence (ECL) sensing device based on a boron carbon nitride/gold nanoparticle (BCN/AuNPs) complex is developed to determine BNP. Prominently, the CRISPR/CAS 13a enzyme reverse cleavage mode, the entropy-driven and hammer hybridization chain reaction processes were involved in the entire detection scheme. Ultimately, with multiple reaction methods and amplification reactions of nucleic acids, this ECL sensing device is able to achieve a detection limit as low as of 0.03 pg/mL and linear range from 0.1 pg/mL to 30 ng/mL for BNP. In addition, the ECL sensing device based on BCN/AuNPs complex obtained satisfactory stability and specificity, and can also be extended to the detection of other pathological markers.
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PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Lingsong Lu,Bei Liu
3D porous Fe 3 O 4 @ZIF-90 interface provides high-density DNA loading sites. • Mn 2+ -gated DNAzyme walker ensures a low-background controllable assay. • DNAzyme walker and CRISPR-Cas13a cascade enable dual signal amplification. • The biosensor demonstrated high accuracy and specificity in human serum samples. The precise quantification of Mucin-1 (MUC1) is vital for the early diagnosis and therapeutic monitoring of epithelial malignancies. In this study, a fluorescent biosensing platform based on a 3D porous metal-organic framework (MOF) and a cascaded signal amplification strategy was developed for MUC1 detection, driven by a target-induced conformational switch and Mn 2+ -dependent catalytic DNAzyme walking. Porous Fe 3 O 4 @ZIF-90 nanocomposites were used to construct the sensing interface, which co-immobilized dormant DNAzyme/aptamer duplexes and a high-density layer of substrate DNA (sub-DNA), serving as a robust platform for the subsequent walking process. Upon specific MUC1 recognition, the aptamer preferentially captures the target MUC1. The thermodynamic driving force induces the dissociation of the MUC1/aptamer complex, liberating the DNAzyme engine. Following magnetic separation to eliminate matrix interferences, the addition of Mn 2+ triggers the processive inter-molecular migration of the DNAzyme walker across the sub-DNA immobilized 3D porous Fe 3 O 4 @ZIF-90 interface via successive hybridization and site-specific cleavage of internal riboadenosine (rA) junctions. This inter-molecular walking process generates a high flux of trigger strands, which subsequently activates the CRISPR-Cas13a system for a robust trans -cleavage of dual-labeled fluorophore-quencher (FQ) reporters. The resulting degradation of FQ reporter enables sensitive MUC1 quantification through exponential fluorescence recovery. By synergizing the structure advantages of the 3D MOF with this cascaded signal amplification, the biosensor achieved a wide linear range from 0.37 nM to 500 nM with a low detection limit of 17.2 pM. Furthermore, the biosensor exhibited reliable specificity and recovery (102.28%–106.94%) in complex human serum assays. The proposed strategy effectively minimizes non-specific background leakage, demonstrating promising potential for the ultra-sensitive detection of MUC1 in complex biological matrices. Schematic presentation of the fluorescent biosensor for the determination of Mucin-1 based on a 3D porous Fe 3 O 4 @ZIF-90 interface and DNAzyme walker-mediated CRISPR-Cas13a cascade. Download: Download high-res image (117KB) Download: Download full-size image
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