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CRISPR-Cas12a with split crRNA for the direct and sensitive detection of microRNA

IF 4.2 3区化学

Analyst Pub Date : 2025-03-19 DOI: 10.1039/d5an00142k

Sijia Yang, Lei Ren, Ningke Fan, Shuhao Wang, Bo Shen, Zhongmei Liu, Xinmin Li, Shijia Ding

{"title":"CRISPR-Cas12a with split crRNA for the direct and sensitive detection of microRNA","authors":"Sijia Yang, Lei Ren, Ningke Fan, Shuhao Wang, Bo Shen, Zhongmei Liu, Xinmin Li, Shijia Ding","doi":"10.1039/d5an00142k","DOIUrl":"https://doi.org/10.1039/d5an00142k","url":null,"abstract":"microRNAs (miRNAs) have been identified as potential biomarkers. Despite the prevalence of quantitative PCR in the field of miRNAs detection, this technology is encumbered by the complexity of its methodology. This study presents a novel CRISPR/Cas12a-based method for direct and sensitive detection of miRNA-21 using split crRNA. The system comprises Cas12a protein, crRNA-handle, and activator DNA complementary to the target miRNA. In the presence of the target miRNA, it binds to the activator DNA, forming a duplex. The formed duplex, in conjunction with the crRNA-handle, activates Cas12a's trans-cleavage activity. This leads to cleavage of a fluorescent reporter, generating an enhanced signal. The method enables direct RNA detection without reverse transcription or sample amplification, offering simplicity and efficiency. This method demonstrates high sensitivity with a minimum detectable limit of 5 pM. Furthermore, the method's specificity is substantiated by its capacity to discern point mutations in miRNA. This system has been shown to quantitatively analyse miRNA-21 levels present within serum, as evidenced by the recovery experiment. Therefore, the method's simplicity, stability, and cost-effectiveness render it a powerful tool for nucleic acid detection, with potential for clinical applications.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"91 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143653985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Effect of precursors on carbon dot functionalization and applications: a review

IF 4.2 3区化学

Analyst Pub Date : 2025-03-19 DOI: 10.1039/d4an01554a

Sara Strickland, Luke Fourroux, Dimitri Pappas

引用次数: 0

Rapid On-site Identification of Pyrrolizidine Alkaloids in Herbal Medicines Using Miniature Mass Spectrometry

IF 4.2 3区化学

Analyst Pub Date : 2025-03-18 DOI: 10.1039/d5an00065c

Xiyue Yang, Xingyu Wang, Yilin Chen, Ziying Wang, Dan Zhang, Nan Wang, Zhengtao Wang, Aizhen Xiong, Linnan Li, Li Yang

{"title":"Rapid On-site Identification of Pyrrolizidine Alkaloids in Herbal Medicines Using Miniature Mass Spectrometry","authors":"Xiyue Yang, Xingyu Wang, Yilin Chen, Ziying Wang, Dan Zhang, Nan Wang, Zhengtao Wang, Aizhen Xiong, Linnan Li, Li Yang","doi":"10.1039/d5an00065c","DOIUrl":"https://doi.org/10.1039/d5an00065c","url":null,"abstract":"Pyrrolizidine alkaloids (PAs) are naturally occurring plant toxins with significant multi-organ toxicity, especially hepatotoxicity. Accidental consumption of PAs-containing herbal medicines can lead to severe health consequences, emphasizing the need for rapid and effective detection methods to ensure medicinal safety. In this study, we developed a novel on-site rapid analytical method using paper capillary spray miniature mass spectrometry (PCS-mini MS), and created a database containing 34 different PAs with a detection limit ranging from 0.5 to 2 ngmL-1. This method is particularly suitable for identifying PAs in herbal medicines, The accuracy of PCS-mini MS was validated through high-performance liquid chromatography-mass spectrometry. Furthermore, the method's environmental impact was assessed using three green evaluation tools, demonstrating its compliance with green analytical chemistry principles, highlighting both efficiency and sustainability. This study provides a convenient and precise approach for regulating the herbal medicine market, enabling quick identification of toxic plants and reducing the risk of adverse health effects from herb misidentification. In the future, this method is expected to be widely adopted for clinical applications and market regulation of herbal products.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"183 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143640415","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Feature selection of Raman spectra for forensic document examination using machine learning

IF 4.2 3区化学

Analyst Pub Date : 2025-03-18 DOI: 10.1039/d4an01529k

Yong Ju Lee, Chang Woo Jeong, Kim Hong Taek, Tai-Ju Lee, Hyoung Jin Kim

{"title":"Feature selection of Raman spectra for forensic document examination using machine learning","authors":"Yong Ju Lee, Chang Woo Jeong, Kim Hong Taek, Tai-Ju Lee, Hyoung Jin Kim","doi":"10.1039/d4an01529k","DOIUrl":"https://doi.org/10.1039/d4an01529k","url":null,"abstract":"Forensics relies on the differentiation and classification of document papers, particularly in cases involving document forgery and fraud. In this study, document paper is classified by integrating Raman spectroscopy with machine learning models, namely, random forest (RF), support vector machines (SVMs), and feed-forward neural networks (FNNs). Among the machine learning models, the RF model effectively caculated the feature importance and identified the critical spectral region contributing to classification, enhancing the transparency and interpretability of the result. Spectral preprocessing with first derivative, significantly improved the classification performance. The spectral range 200–1,650 cm⁻¹ was identified as a highly informative region for differntiation, reducing the number of input variables from 756 to 360 while enhancing the model accuracy. The FNN model outperformed the RF and SVM, with an F1 score of 0.968. The results underscore the potential of combining Raman spectroscopy with machine learning for forensic document examination, offering an interpretable, computationally efficient, and robust approach for paper classification.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"69 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143640417","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A point-of-need framework for illicit drug identification with high-resolution mass spectrometry

IF 4.2 3区化学

Analyst Pub Date : 2025-03-18 DOI: 10.1039/d5an00082c

Thomas P Forbes, Elizabeth L. L Robinson, Edward Sisco, Abigail Koss

{"title":"A point-of-need framework for illicit drug identification with high-resolution mass spectrometry","authors":"Thomas P Forbes, Elizabeth L. L Robinson, Edward Sisco, Abigail Koss","doi":"10.1039/d5an00082c","DOIUrl":"https://doi.org/10.1039/d5an00082c","url":null,"abstract":"The continually evolving drug landscape, with novel synthetic drugs and unique compositions, necessitates the need to advance technologies, data analysis methods, and data accessibility for compound detection and identification. Providing public health, first responder, and law enforcement communities with critical information in near real-time will aid emergency response and public awareness, and direct overdose prevention and interdiction efforts. A major component of this framework is the progression of accurate drug screening and preliminary identifications from a more rigid laboratory-based arrangement to an agile point-of-need paradigm. We investigated drug detection and identification of a field deployable ruggedized high-resolution time-of-flight mass spectrometer, employing both acetone-assisted vacuum ultraviolet (VUV) photoionization and dielectric barrier discharge ionization (DBDI) schemes. This preliminary fit-for-purpose exploration was conducted under laboratory conditions, considering ion sources not reliant on helium gas or external roughing pumps, building toward deployment in a mobile laboratory setting. The chromatography-free measurements enabled rapid analysis of neat drug solutions and multi-component mixtures. Characterization and optimization of system parameters demonstrated sensitive performance, with limits of detection in the tens to hundreds of picograms for a range of drug classes from multiple-component mixtures. The system’s high mass resolution was calibrated with a polyethylene glycol calibrant, enabling accurate matching with spectral library entries. Integrating compound identification with the NIST DART-MS Forensics Database and NIST/NIJ DART-MS Data Interpretation Tool provided a solid foundation for transition to the point-of-need. The overarching framework seeks to support technology advancement and adoption, as well as the development of novel data analysis tools, processes, and management for public access and utilization.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"23 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143640414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tuning FeⅡ/FeⅢ ratio by substituent regulation to enhance peroxidase-like activity of metal–organic framework for sensitive biosensing

IF 4.2 3区化学

Analyst Pub Date : 2025-03-15 DOI: 10.1039/d4an01448k

Keyu Xing, Siyao Peng, Mengting Liu, Ling-Li Long, Shao-Yi Yu, Li Yao, Jian Zhang, Mao-Long Chen, Yun-Hui Cheng, Zhou Xu

引用次数: 0

CRISPR/Cas12a-Powered Nanoconfined Biosensing Platform with Hybrid Chain Reaction Cascading Guanine Nanowire Amplification for Ultrasensitive Dual-Mode Detection of Lipopolysaccharide

IF 4.2 3区化学

Analyst Pub Date : 2025-03-14 DOI: 10.1039/d5an00184f

Yanlei Li, Xiang Ren, Dan Wu, Hongmin Ma, Qin Wei, Huangxian Ju, Zhongfeng Gao

{"title":"CRISPR/Cas12a-Powered Nanoconfined Biosensing Platform with Hybrid Chain Reaction Cascading Guanine Nanowire Amplification for Ultrasensitive Dual-Mode Detection of Lipopolysaccharide","authors":"Yanlei Li, Xiang Ren, Dan Wu, Hongmin Ma, Qin Wei, Huangxian Ju, Zhongfeng Gao","doi":"10.1039/d5an00184f","DOIUrl":"https://doi.org/10.1039/d5an00184f","url":null,"abstract":"Traditional endotoxin detection methods face challenges in sensitivity, interference resistance, and reliability. This study develops a CRISPR/Cas12a-powered nanoconfined biosensing system that integrates mesoporous nanoengineering with a hybrid chain reaction (HCR) cascading guanine nanowire (G-wire) dual amplification strategy for ultrasensitive dual-mode detection of lipopolysaccharide (LPS). By leveraging a vertically ordered mesoporous silica membrane (VMSM) as a molecular sieve and CRISPR trans-cleavage activity modulator, the system achieves precise regulation of Ru(bpy)₃²⁺ adsorption via LPS-suppressed HCR assembly. This architecture enables physical confinement-mediated electrochemiluminescence (ECL) and fluorescence (FL) signal transduction, with dual-mode outputs providing mutual validation for enhanced reliability. The biosensor exhibits superior sensitivity with detection limits of 3.4 pg/mL for ECL and 1.4 pg/mL for FL, while also offering a broad dynamic range (0.005–100 ng/mL), significantly outperforming conventional LPS assays. The CRISPR-triggered HCR cascading G-wire dual amplification synergizes with nanoconfinement of VMSM to ensure robust anti-interference performance in complex matrices, validated by recovery rates of 97.8–102.5% in real samples. By integrating CRISPR programmability with nanoengineered signal amplification, this work establishes a transformative paradigm for portable, high-precision endotoxin detection in clinical diagnostics, industrial monitoring, and environmental safety applications.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"41 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143618697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Paper-based visual colorimetric platform for rapid detection of arsenic in the environments

IF 4.2 3区化学

Analyst Pub Date : 2025-03-14 DOI: 10.1039/d5an00131e

Zhiqiang Wang, Feng He, Xinxin Xu, Hua Kuang, Chuanlai Xu

引用次数: 0

Raman Spectroscopy as a Comprehensive Tool for Profiling Endospore-forming Bacteria

IF 4.2 3区化学

Analyst Pub Date : 2025-03-13 DOI: 10.1039/d5an00115c

Markus Salbreiter, Petra Rösch, Annette Wagenhaus, Juergen Popp

{"title":"Raman Spectroscopy as a Comprehensive Tool for Profiling Endospore-forming Bacteria","authors":"Markus Salbreiter, Petra Rösch, Annette Wagenhaus, Juergen Popp","doi":"10.1039/d5an00115c","DOIUrl":"https://doi.org/10.1039/d5an00115c","url":null,"abstract":"Accurate and reliable bacterial identification at the genus and species levels is essential for effective clinical diagnostics. Pathogens such as Clostridium perfringens, Bacillus cereus, Clostridioides difficile, and Paraclostridium sordellii pose significant challenges due to their unique cultivation requirements and developmental traits. Building on our previous work demonstrating the differentiation of vegetative Clostridium cells from non-Clostridium genera, we now aim to extend this approach to distinguish endospores of the same species. Raman spectroscopy was utilized to develop a comprehensive library of endospore spectra, encompassing both pathogenic and non-pathogenic species. This extensive dataset forms the foundation for advanced analytical capabilities. Chemometric analysis of single-endospore Raman spectra revealed significant discriminatory power across multiple hierarchical levels, facilitating the distinction between vegetative cells and endospores. Furthermore, this method enabled precise genus- and species-level classification of endospores, underscoring its potential for high-resolution bacterial endospore identification. These findings highlight the versatility and efficacy of Raman spectroscopy in addressing the challenges associated with the identification of bacterial endospores in diverse clinical and environmental contexts. These findings present the first comprehensive library of endospore Raman spectra, demonstrating that Raman spectroscopy combined with chemometric analysis is a robust and reliable method for differentiating endospores of Clostridium species from those of Bacillus, Clostridioides, and Paraclostridium. This approach holds significant promise as a precise diagnostic tool for bacterial endospore identification in clinical settings.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"30 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608113","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

One-pot synthesized three-way junction based multiple strand displacement amplification for sensitive assay of H5N1 DNA

IF 4.2 3区化学

Analyst Pub Date : 2025-03-13 DOI: 10.1039/d4an01586j

Zhengjiang Wu, Jingwen Li, Tao Zhang, Kai Zhang, Xiaomei Liu, Zhan Yang, Li Xu, Kun Han

{"title":"One-pot synthesized three-way junction based multiple strand displacement amplification for sensitive assay of H5N1 DNA","authors":"Zhengjiang Wu, Jingwen Li, Tao Zhang, Kai Zhang, Xiaomei Liu, Zhan Yang, Li Xu, Kun Han","doi":"10.1039/d4an01586j","DOIUrl":"https://doi.org/10.1039/d4an01586j","url":null,"abstract":"The rapid and sensitive detection of H5N1, a highly pathogenic avian influenza virus, is crucial for controlling its spread and minimizing its impact on public health. In this study, we developed a novel biosensor based on strand displacement amplification (SDA) coupled with CRISPR/Cas12a for highly sensitive detection of H5N1 DNA. The biosensor utilizes a combination of a three-way junction structure, composed of three hairpins (H1, H2, H3), to initiate amplification through SDA, resulting in the production of numerous activators. These activators then trigger CRISPR/Cas12a's collateral cleavage activity, which generates a detectable fluorescence signal. The biosensor demonstrated a linear detection range from 100 fM to 800 pM, with a detection limit as low as 72.87 fM. The optimized biosensor exhibited excellent sensitivity, high specificity, and a broad dynamic range, making it a promising tool for the early detection of H5N1 DNA in complex biological samples. Additionally, the use of CRISPR/Cas12a's trans-cleavage activity significantly improved signal amplification and specificity, allowing for more reliable detection compared to traditional methods. The results highlight the advantages of the integrated SDA and CRISPR/Cas12a approach, which addresses the limitations of conventional detection methods, such as low sensitivity, lengthy analysis times, and high costs. The biosensor's ability to perform well in complex sample matrices demonstrates its potential for point-of-care diagnostics, especially in resource-limited settings. Future applications of this technology could extend to the detection of other pathogens, offering a versatile and adaptable platform for disease surveillance and management.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"14 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143608498","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0