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.
研究领域:
生物传感器
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病毒学
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纳米技术
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分子诊断学
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CRISPR技术
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分析化学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Chemistry
·
2025
·
IF 4.2
Ting Yuan,Jianbo Yuan,Jian Huang,Nana Li,Huan Cai,Yan Yang,Juan Li,Rui Chen,Xun Min
Introduction: The rapid and accurate identification of Mycobacterium tuberculosis (MTB) is essential for effective tuberculosis (TB) control. However, conventional diagnostic methods for MTB suffer from limitations such as low sensitivity, poor specificity, high cost, reliance on specialized instruments, and complex, time-consuming procedures. To address these challenges, there is an urgent need for a simple, rapid, and highly sensitive detection method that can be deployed in point-of-care settings.Methods: We developed a one-pot biosensing system combining recombinase polymerase amplification (RPA) and CRISPR/Cas12a-driven G4-hemin self-assembly for the colorimetric detection of MTB. Glycerol was employed as a phase-separation barrier to prevent interference between RPA amplification and CRISPR/Cas12a trans-cleavage. A single-stranded DNA (ssDNA) probe, designed to self-assemble with ssDNA-hemin into G4-hemin nanozymes upon CRISPR/Cas12a-mediated cleavage, served as the reaction substrate. The ssDNA-hemin further enhanced the catalytic activity of the generated G4-hemin DNAzyme. The entire assay was completed in a single step within 60 min without requiring complex instrumentation.Results and Discussion: Under optimized conditions, the biosensing system achieved ultrasensitive naked-eye detection of MTB with a limit of detection (LOD) of 10 copies/μL, comparable to traditional four-step fluorescent assays. Clinical validation using 104 patient samples demonstrated high concordance with standard diagnostic methods. This approach combines the advantages of recombinase polymerase amplification (RPA), CRISPR/Cas12a specificity, and G4-hemin DNAzyme-based colorimetric signal amplification, enabling simple, equipment-free visual detection. Given its speed, sensitivity, and ease of use, this biosensing system holds significant promise for point-of-care MTB nucleic acid testing in resource-limited settings.
研究领域:
生物传感器
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分子诊断
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CRISPR技术
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核酸扩增
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DNA酶技术
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结核病检测
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2025
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IF 6
Hong Zhang,Xuekun Bai,Chenxi Wang,Wei Pang,Jiaxin Zhang,Junhuai Song,Huanying Zhou,Zhixian Gao,Bin Zheng
Background Biorhythmic played an important role in physical health. Biorhythmic disorders led to various health issue, including cardiovascular diseases and metabolic disorders. Early diagnosis of biorhythmic disorders was crucial in slowing the progression of disease. Therefore, it was important to develop technologies for detecting biorhythmic and their disorder markers. This study selected HMGB1(High mobility group box 1), BMAL1 (Brain and Muscle Aryl hydrocarbon receptor nuclear translocator-like 1), and MICU (mitochondrial calcium uptake 1) mRNA for detection. Results CRISPR/Cas12a/SERS (surface-enhanced Raman spectroscopy) ultrasensitive biosensing platform was developed for the detection of biorhythmic markers. The biosensor was partly an ultrabright Raman signal reporter (on) of a novel satellite structure formed by gold/silver bimetallic nanoparticles. This novel satellite structure utilizes the potential difference between gold and silver nanoparticles to promote the transfer of electrons from the core structure to the AuNPs and MBA (4-mercaptobenzoic acid) molecules, while enhancing the electromagnetic field distribution. Compared with the Raman signal of the satellite structure composed of Au/Au bimetallic, the Raman signal of the satellite structure in this study is increased by about 10 times. CRISPR/Cas12a recognized the target molecule and activated trans -cleavage activity, cleaving the DNA/RNA hairpin structure and releasing RNA 2∗ . RNA 2∗ caused the satellite structure to disperse, significantly attenuating the ultra-strong Raman signal (off), thereby enabling quantitative detection of the target nucleic acid. The LOD for three targets were of less than 1 fM. Significance In summary, the CRISPR/Cas12a/SERS biosensing platform enables mRNA detection without preamplification. This biosensing platform can detect target molecules at the fM level, achieving ultra-sensitive detection. In addition, the satellite structure exhibits long-term stability, providing a theoretical basis for the practical application of this biosensing platform.
研究领域:
生物传感器
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分子诊断学
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纳米技术
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CRISPR技术
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RNA检测
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
ACS Sensors
·
2025
·
IF 9.1
Lingxuan Qu,Lingyi Meng,Xilin Sun,Wei Cui,Jin Shi,Yuqi Wang,Qingshuang Hu,Shuping Xu,Bingxin Sun,Chongyang Liang
Aiming at realizing field detection during HIV-1 patient screening in developing countries and point of care testing (POCT) of virus levels in HIV-1 patients receiving medication, an urgent demand for portable nucleic acid detection technology with low cost and sensitivity is raised. To solve this, a CRISPR/Cas13a-based electrochemical detection platform by a multiple combined crRNAs strategy is developed. This sensing platform is based on the Ion Current Rectification regulation through a Porous Anodic Alumina membrane decorated with ssRNA chains, which are trans-cleavage substrates for activated Cas13a and become shorter when the target gene exists, resulting in an altered motion of transmembrane ions. This platform was proven to be feasible and ultrasensitive for tracing the HIV-1 virus by multiple applicable crRNAs combination, and its clinical sample detection limit arrives at 60 copies/μL for HIV-1 patients’ serum samples. Importantly, this method is amplification-free, and its sensitivity is beyond the nonamplification fluorescence-based nucleic acid strategy by one order of magnitude. This sensor is facile to use and cost-effective, and can also be extended to other RNA-based pathogenic microorganism detections. This POCT method facilitates self-monitoring of drug resistance in HIV-1 patients, regardless of medication status.
研究领域:
生物传感器
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分子诊断
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病毒学
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生物电子学
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即时检测
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CRISPR技术
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纳米技术
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RNA检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2025
·
IF 10.5
Junlan Liu,Yuan Gao
The exceptionally sensitive and specific CRISPR-Cas systems have revolutionized molecular diagnostics, but their application remains largely confined to nucleic acid targets. To overcome this limitation and enable one-pot, mix-and-read detection of non-nucleic acid analytes, we present a generalizable CRISPR-Cas13a sensing platform that utilizes a hybrid DNA:RNA strand displacement-based switch to conditionally activate Cas13a. The switch leverages Cas13a's strict requirement for single-stranded RNA activators to mediate non-RNA target-triggered activation while protecting functional DNA elements from collateral cleavage. By integrating the modular Cas13a switch with various functional DNAs, including DNAzyme, aptazyme, and duplexed aptamer, we constructed a suite of one-pot sensors capable of detecting non-RNA targets such as lead ion, ATP, and thrombin. These CRISPR-Cas13a sensors demonstrate enhanced sensitivity, high selectivity, and robust performance in complex biological matrices. The high modularity of the CRISPR-Cas13a sensor allows rapid reconfiguration for new targets by simply altering the output sequence of functional DNA without extensive re-engineering. This versatile and robust sensing strategy expands the application landscape of CRISPR-Cas systems and lays the foundation for flexible, sensitive, and one-pot detection of a wide array of clinically and environmentally relevant targets.
研究领域:
分子诊断学
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合成生物学
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生物传感器
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CRISPR技术
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核酸化学
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生物分析化学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Qiyu Zhang,Zhaoying Liu,Enze Sheng,Zhenjiang Liu,Rujin Zhao,Ming Li
To address the challenge of readily available and cost-effective monitoring, a novel aptasensor platform was proposed relying on the clustered regularly interspaced short palindromic repeats/associated protein 12a (CRISPR/Cas12a)-driven and glucosemeter-assisted system for zearalenone (ZEN). The magnetic bead (MB)-based probes of MB@Apt-cDNA and MB@invertase were prepared to provide specific recognition and efficient conversion. When ZEN presented, the released cDNA activated the cutting function of CRISPR/Cas12a for ssDNA, releasing numerous invertase to convert the environmentally friendly sucrose. Then, the generated glucose signals were positively related to ZEN levels, which quantified by a glucosemeter. The proposed aptasensor displayed an excellent sensitivity, achieving the limit of detection (LOD) and detection range of 0.218 ng/mL and 0.218−109.89 ng/mL, respectively. The platform also exhibited high specificity (with cross-reactivity values below 6.81 % for other analytes) and high accuracy for ZEN detection. The satisfactory reliability and practicability had been demonstrated through its application in real agricultural byproduct samples. The area under the curve was 0.979, with an optimal operating point being (0.125, 1.000). This study can provide an alternative pathway for improving the detection performance and accessibility to public in limited situation, suggesting the efficient CRISPR/Cas12a and green glucosemeter detection strategies for other pollutant monitoring.
研究领域:
生物传感器
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分析化学
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食品安全
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分子诊断
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环境科学
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CRISPR技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
METHODS
·
2025
·
IF 4.3
Wensi Li,Zhichao Tang,Xiang Zhu,Beijuan Wang,Xiuqin Zhang,Ze Ji,Suhang Cheng
In this work, an electrochemiluminescence (ECL) biosensor integrating T7 RNA polymerase amplification and CRISPR/Cas13a-mediated signal enhancement was developed for the ultrasensitive detection of matrix metalloproteinase-2 (MMP-2), a key biomarker associated with chronic inflammatory diseases such as COPD. A peptide nucleic acid (PNA) probe was designed to respond specifically to MMP-2 cleavage, enabling the release of DNA templates for subsequent T7 RNA polymerase-driven transcription amplification. The generated RNA triggers the collateral cleavage activity of CRISPR/Cas13a, resulting in a significant amplification of the ECL signal. The biosensor’s surface was constructed using a AuNPs/Ti 3 C 2 Tx/Ru(II)-PEI nanocomposite, which enhanced signal transduction and stability. Under optimized conditions, the proposed biosensor achieved a detection limit as low as 62.05 fM, demonstrating superior sensitivity compared to conventional methods, as summarized in Table 1 . The platform also exhibited excellent specificity and anti-interference capability, ensuring reliable detection of MMP-2 in complex biological samples. This study provides a simple yet highly efficient strategy for enzymatic biomarker detection, offering great potential for clinical applications in early disease diagnosis and monitoring.
研究领域:
生物传感器
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分子诊断
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CRISPR技术
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纳米技术
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慢性阻塞性肺疾病
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酶检测
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Frontiers in Microbiology
·
2025
·
IF 4.5
Yayun Jiang,Zongyao Chen,Xiao Liu,Qi Xin,Dengchao Wang,Caixia Ji,Youwei Li,Gang Mai
Methicillin-resistant Staphylococcus aureus (MRSA), a widely drug-resistant bacterium, poses a significant threat to global health. Current culturing and nucleic acid detection methods are time-consuming and require complex instruments, which do not meet the detection needs. Herein, we developed a rapid and visual MRSA detection method (MCFHCR) using ssDNA-functionalized magnetic beads as a trigger chain combined with the trans-cleavage activity of the Cas12a protein and fluorescence signal amplification of the hybridization chain reaction (HCR). MCFHCR is a signal-off platform for the detection of MRSA. In the absence of DNA targets, the trans-cleavage activity of Cas12a is inactivated, allowing HCR to proceed and form long double-stranded DNA, resulting in an increased fluorescent signal. In the presence of the DNA targets, the trans-cleavage activity of CRISPR/Cas12a is activated to cleave the trigger strand, failing HCR and leading to a decrease in the fluorescence signal. Combined with RPA, MCFHCR was completed within 35 min, achieving a limit of detection (LOD) of five copies/μL for mecA DNA and 8 CFU/mL for MRSA. In detecting clinical strains, MCFHCR demonstrated comparable performance to qPCR and drug sensitivity testing. Therefore, with its simple, rapid operation and convenient signal acquisition, MCFHCR shows significant practical applicability in detecting MRSA.
研究领域:
微生物学
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分子诊断学
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CRISPR技术
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传染病检测
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生物传感器
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2025
·
IF 4.6
Anjie Guo,Yu Wu,Yunkai Xie,Wang Guo,Yiqing Guo,Xinai Zhang,Wen Zhang,Xiaobo Zou,Zongbao Sun
Di-(2-ethylhexyl) phthalate (DEHP) can leach into food and the environment, posing health and ecological risks. This paper introduces a novel CRISPR-based fluorescent aptasensor for on-site DEHP detection. The aptasensor selectively binds to DEHP, triggering a competitive displacement reaction that releases an Aptamer-dsDNA probe. After magnetic separation, the supernatant is analyzed via a Cas12a reporter system, where Cas12a activation cleaves a FAM-BHQ reporter, generating fluorescence. Combined with smartphone imaging, this method enables rapid result acquisition. The aptasensor shows high selectivity and sensitivity, detecting DEHP from 1 pg/mL to 1 μg/mL, with a lower limit of 0.15 pg/mL. It effectively detects DEHP in various real samples, offering reliable visual monitoring. This method offers a rapid and effective on-site detection strategy for food safety and environmental pollution monitoring.
研究领域:
食品安全
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环境监测
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生物传感器
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分子诊断
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CRISPR技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
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
·
CRISPR技术
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即时检测
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传染病检测
✓ 已验证
产品关联
DOI
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Rui Qiu,Xiaolan Lin,Canyan Yang,Xian Chen
Vancomycin is a natural glycopeptide antibiotic produced by Streptomyces species. It is primarily employed to treat infections caused by drug-resistant bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA). However, the therapeutic concentration range of vancomycin is narrow, necessitating therapeutic drug monitoring (TDM). Currently, the commonly used vancomycin detection methods face challenges such as high equipment costs, complex operations, lengthy processing times, and poor reproducibility. To overcome these limitations, we developed a CRISPR-enhanced fluorescent (CEF) aptasensor for efficient and accurate detection of vancomycin, using signal amplification to provide critical support for TDM. In the presence of vancomycin, its specific binding to the aptamer exposes the primer strand, which is part of the self-assembled DNA tetrahedron, thereby initiating the RCA process. The product subsequently activates Cas12a, causing it to trans-cleave the signal probe ssDNA-FQ, which is modified by Cy3 and BHQ2. The fluorophore will move away from the quencher, thereby restoring fluorescence and generating a significantly amplified fluorescent signal, achieving highly sensitive detection of vancomycin. The constructed biosensor exhibits good linear response in the target concentration range of 1 fM to 10 nM, with a detection limit of 164 aM. This novel biosensor holds great promise for the accurate detection of low-abundance biomarkers like vancomycin, which is crucial for ensuring optimal dosing and preventing adverse effects such as nephrotoxicity. Moreover, the simplicity, rapidity, and cost-effectiveness of this aptasensor make it an ideal candidate for therapeutic drug monitoring and clinical diagnostics.
研究领域:
生物传感器
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分子诊断
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抗生素监测
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CRISPR技术
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分析化学
✓ 已验证
产品关联
DOI
高影响因子
Cell Reports
·
2023
·
IF 8.8
Ting Zeng,Jie Yin,Ziwen Liu,Zhaoxing Li,Yu Zhang,Yang Lv,Mei-Ling Lu,Min Luo,Meirong Chen,Yibei Xiao
Summary The type V-K CRISPR-associated transposons (CASTs) allow RNA-guided DNA integration and have great potential as a programmable site-specific gene insertion tool. Although all core components have been independently characterized structurally, the mechanism of how the transposase TnsB associates with AAA+ ATPase TnsC and catalyzes donor DNA cleavage and integration remains ambiguous. In this study, we demonstrate that TniQ-dCas9 fusion can direct site-specific transposition by TnsB/TnsC in ShCAST. TnsB is a 3′-5′ exonuclease that specifically cleaves donor DNA at the end of the terminal repeats and integrates the left end prior to the right end. The nucleotide preference and the cleavage site of TnsB are markedly different from those of the well-documented MuA. We also find that TnsB/TnsC association is enhanced in a half-integration state. Overall, our results provide valuable insights into the mechanism and application expansion of CRISPR-mediated site-specific transposition by TnsB/TnsC.
研究领域:
分子生物学
·
基因工程
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CRISPR技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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