Analyst最新文献_第6页

Reduction of Background-Triggered Amplification in Lesion-Induced DNA Amplification (LIDA)

IF 4.2 3区化学

Analyst Pub Date : 2025-03-06 DOI: 10.1039/d5an00047e

Anantha Srujana Ealeswarapu, Nahida Akter, Julianne Gibbs

{"title":"Reduction of Background-Triggered Amplification in Lesion-Induced DNA Amplification (LIDA)","authors":"Anantha Srujana Ealeswarapu, Nahida Akter, Julianne Gibbs","doi":"10.1039/d5an00047e","DOIUrl":"https://doi.org/10.1039/d5an00047e","url":null,"abstract":"Lesion-induced DNA amplification (LIDA) enables isothermal amplification of nucleic acids, and the only enzyme required is T4 DNA ligase. However, the application of LIDA for the amplification of trace amounts of nucleic acids has been hindered by the observed background-triggered amplification in the absence of initial target due to a pseudo-blunt end ligation reaction of two of the primers. In this work, we have tested three approaches to minimize the background-triggered amplification: increasing and decreasing the concentration of salts such as NaCl and MgCl2, respectively, and increasing the concentration of ATP. All these optimizations sharply decreased the background-triggered amplification. Employing the most favourable buffer condition of 2.5 mM MgCl2 where the target-initiated amplification was least affected while reducing the background-triggered process enabled us to achieve a detection range of 14 nM-140 aM with an approximate limit of detection of 680 aM, which is six orders of magnitude more sensitive than using our standard amplification conditions. This optimization of the salt and co-factor concentrations to decrease background and enhance the sensitivity of LIDA has demonstrated LIDA’s potential for application in clinical diagnostics.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"16 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143570027","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

Copper doped NH2-metal-organic frameworks as co-reactant modulating unit for sensitive electrochemiluminescence immunoassay

IF 4.2 3区化学

Analyst Pub Date : 2025-03-06 DOI: 10.1039/d5an00032g

Shuai-Fei Zhang, Yu-Ling Wang, Xiao-Long Fu, Shu-Wei Ren, Juntao Cao, Yan-Ming Liu

{"title":"Copper doped NH2-metal-organic frameworks as co-reactant modulating unit for sensitive electrochemiluminescence immunoassay","authors":"Shuai-Fei Zhang, Yu-Ling Wang, Xiao-Long Fu, Shu-Wei Ren, Juntao Cao, Yan-Ming Liu","doi":"10.1039/d5an00032g","DOIUrl":"https://doi.org/10.1039/d5an00032g","url":null,"abstract":"A facile electrochemiluminescence (ECL) immunosensor for the sensitive detection of prostate specific antigen (PSA) was constructed based on a co-reactant modulating strategy using Cu2+-modified NH2-metal-organic frameworks (NMOF@Cu2+) nanoprobes as modulating units. In this proposal, the biconical NMOF@Cu2+ nanoprobes possess a large number of carboxyl groups, which could be labeled by the targets PSA to form PSA-NMOF@Cu2+ conjugates. Moreover, the nanoprobes could consume K3[Fe(CN)6] and result in a lower signal of the ECL system of luminol-K3[Fe(CN)6]. The bioassay was executed in a split-type mode. First, the competitive immunological recognition reaction with PSA-NMOF@Cu2+ conjugates as the signal probes were performed in the antibody labeled 96-well plate. Upon the targets PSA were present, PSA will compete with the PSA-NMOF@Cu2+ conjugates for immobilization, which determined the amount of PSA-NMOF@Cu2+ conjugates immobilized in the plate and resulted in different amounts of the prefilled K3[Fe(CN)6] being consumed. Then, the above reacted solution was transferred in the detection cell for ECL test, the ECL signal from luminol/K3[Fe(CN)6] system will reflect the content of PSA. The developed ECL immunosensor showed high sensitivity for PSA with a linear response range of 1.0 pg·mL−1 ‒ 10 ng·mL−1 and a detection limit of 0.5 pg·mL−1. Moreover, the applicability of the present method was demonstrated in the determination of PSA in human serum.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"191 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143561198","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

Nicking enzyme assisted amplification combined with CRISPR-Cas12a system for one-pot sensitive detection of APE1†

IF 3.6 3区化学

Analyst Pub Date : 2025-03-05 DOI: 10.1039/D5AN00009B

Wei Dai, Han Wang, Hanxu Ji, Xian Xiao, Yiyuan Li, Dayang Jiang, Yangkang Luo, Xianjin Xiao, Bei Yan, Jie Yu and Longjie Li

{"title":"Nicking enzyme assisted amplification combined with CRISPR-Cas12a system for one-pot sensitive detection of APE1†","authors":"Wei Dai, Han Wang, Hanxu Ji, Xian Xiao, Yiyuan Li, Dayang Jiang, Yangkang Luo, Xianjin Xiao, Bei Yan, Jie Yu and Longjie Li","doi":"10.1039/D5AN00009B","DOIUrl":"10.1039/D5AN00009B","url":null,"abstract":"Apurinic/apyrimidinic endonuclease 1 (APE1) is a critical enzyme in the base excision repair (BER) pathway, essential for preserving cellular equilibrium. Variations in APE1 activity within blood or tissues can provide significant insights for clinical cancer screening and disease diagnosis. Consequently, the detection of APE1 activity is critical for clinical diagnostics. However, there is currently a deficiency in rapid, straightforward, and sensitive methods for APE1 detection. To address this issue, we developed a method that integrates Nicking Enzyme Assisted Amplification (NEAA) with CRISPR-Cas12a signal amplification, enabling one-pot detection of APE1 activity. This method utilizes NEAA to produce a substantial quantity of target DNA that is complementary to the crRNA, thereby triggering the trans-cleavage activity of Cas12a. The activated Cas12a then amplifies and emits signals by cleaving the reporter probe. Our strategy allows for the swift and precise detection of APE1, in only 3 h, with a detection threshold of 1 × 10−6 U mL−1 and a linear detection range of 5 × 10−6 to 0.1 U mL−1. It has been effectively utilized for the detection of APE1 in biological samples.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1409-1418"},"PeriodicalIF":3.6,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143546982","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

The fabrication of 1,2-dicarbonyl compound-caging isothermal exponential amplification and its application in highly sensitive detection of tumor exosomal miRNA

IF 4.2 3区化学

Analyst Pub Date : 2025-03-04 DOI: 10.1039/d4an01515k

Huiyu Pan, Jiahui Dong, Xuyang Shen, Lingli Zhao, Ying Liu, Yu-Nan Chen, Zi-Qin Huang, Ying-Lin Zhou, Xinxiang Zhang

{"title":"The fabrication of 1,2-dicarbonyl compound-caging isothermal exponential amplification and its application in highly sensitive detection of tumor exosomal miRNA","authors":"Huiyu Pan, Jiahui Dong, Xuyang Shen, Lingli Zhao, Ying Liu, Yu-Nan Chen, Zi-Qin Huang, Ying-Lin Zhou, Xinxiang Zhang","doi":"10.1039/d4an01515k","DOIUrl":"https://doi.org/10.1039/d4an01515k","url":null,"abstract":"The exponential amplification reaction (EXPAR) possesses the advantage of high amplification efficiency of producing large amounts of short nucleic acids in a short time. However, primer-independent DNA autosynthesis and ab initio DNA self-synthesis mediated nonspecific amplification increase the risk of high background signal, which limits its application in the fabrication of ultrasensitive sensors. Here, we developed a new strategy called 1,2-dicarbonyl compound-caging EXPAR (caging-EXPAR) by innovatively modifying EXPAR templates with 1,2-dicarbonyl compounds. Five 1,2-dicarbonyl compounds including glyoxal and ninhydrin were chosen, all of which were proved to specifically bind to guanosine on EXPAR template, efficiently reducing the background amplification of EXPAR and exhibiting an obviously improved discrimination between target and background. The possible mechanisms of the role of 1,2-dicarbonyl compounds played in EXPAR were proposed, and three possible factors which might induce the serious nonspecific amplification were investigated and verified. Caging-EXPAR provided a general, simple, convenient and beneficial strategy to slow down the generation of EXPAR background signals. Based on this method, choosing miRNA let-7a as a model, the minimum detectable amount was 10 zmol, which was reduced by 3 orders of magnitude compared to traditional EXPAR. More importantly, the strategy was successfully applied to monitor the expression level of the low-abundant miRNA in MCF-7 cell derived exosomes. This highly portable and cost-effective method can not only promote the real-time quantitative detection of specific nucleic acids in many fields such as clinical diagnosis and prognostic treatment, but also provide a complementary theoretical support for EXPAR background amplification, which is conducive to the improvement and wide application of other amplification reactions.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"16 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143539173","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 microfluidic device for serum zinc assay by colorimetry†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-28 DOI: 10.1039/D5AN00023H

Kalpita Nath, Debasish Sarkar and Sunando DasGupta

{"title":"Paper-based microfluidic device for serum zinc assay by colorimetry†","authors":"Kalpita Nath, Debasish Sarkar and Sunando DasGupta","doi":"10.1039/D5AN00023H","DOIUrl":"10.1039/D5AN00023H","url":null,"abstract":"Zinc is an essential micronutrient playing several crucial roles in human pathophysiology and its deficiency leads to micronutrient malnutrition. Therefore, a rapid, inexpensive, and accurate protocol for serum zinc concentration measurement becomes essential in community healthcare. This study demonstrates the design, fabrication, and characterization of a low-cost, paper-based microfluidic device (μPAD) to detect serum zinc concentration by colorimetric techniques. The μPAD comprises circular spotting zones doped with diphenylthiocarbazone, commonly known as dithizone, that produces pink-colored chelates upon reacting with zinc and the color intensity monotonically changes with concentration even across the physiological range (i.e., 5–25 μM). The design and the doping protocol were optimized to generate a linear correlation (in water, R2 = 0.94; in artificial plasma, R2 = 0.98) between a suitable optical measure (i.e., the normalized Euclidean shift) evaluated by image analysis of photographs captured by the camera of a standard smartphone and zinc concentration. The calibration curve for artificial plasma was further used to evaluate the zinc concentrations in real blood serum samples, resulting in a high parity with the respective gold standard method. The device is expected to significantly contribute in diagnosis of micronutrient malnutrition with a particular emphasis on community healthcare and to reach resource-limited settings.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1347-1360"},"PeriodicalIF":3.6,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143518285","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

Extracellular vesicles derived from ovarian cancer cell lines discriminated by biochemical and Fourier transform infrared spectroscopy approaches†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-28 DOI: 10.1039/D5AN00024F

Lefkothea Pantazi, Valérie Untereiner, Paolo Rosales, Romain Rivet, Sandra Audonnet, Isabelle Proult, Laurent Ramont, Ganesh D. Sockalingum and Stéphane Brézillon

{"title":"Extracellular vesicles derived from ovarian cancer cell lines discriminated by biochemical and Fourier transform infrared spectroscopy approaches†","authors":"Lefkothea Pantazi, Valérie Untereiner, Paolo Rosales, Romain Rivet, Sandra Audonnet, Isabelle Proult, Laurent Ramont, Ganesh D. Sockalingum and Stéphane Brézillon","doi":"10.1039/D5AN00024F","DOIUrl":"10.1039/D5AN00024F","url":null,"abstract":"Ovarian cancer is the most lethal cancer among gynaecological malignancies. Due to the lack of early symptoms and screening tools, patients are diagnosed in advanced stages. Cancer invasion and metastasis through the extracellular matrix (ECM) are enhanced by tumour cell Extracellular Vesicles (EV). The aim of this study was to characterise the EVs derived from two ovarian cancer cell lines (ES2 and SKOV3) using biochemical and vibrational spectroscopic approaches. EVs were prepared by ultracentrifugation and characterised by Nanoparticle Tracking Analysis. Their surface proteins were assessed by MACSPlex EV kit for human exosomes. The presence of MMP14 and integrin subunits was evaluated in EVs and cell protein extracts by Western immunoblotting. Both EVs and cells were measured by Fourier transform infrared spectroscopy (FTIR) and data were analysed by hierarchical cluster analysis (HCA). Spectral differences were observed in the lipids and polysaccharides regions both between the SKOV3 and ES2 cells and their corresponding EVs, which allowed a good delineation by HCA. The differences in the biochemical data were confirmed by similar and specific features exhibited in their respective infrared spectral signatures. ES2 EVs exhibited an enrichment in MMP14 in agreement with the aggressiveness of this ovarian cancer metastatic cell line.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1280-1292"},"PeriodicalIF":3.6,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143518286","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

FEN1-assisted LAMP for specific and multiplex detection of pathogens associated with community-acquired pneumonia†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-28 DOI: 10.1039/D4AN01516A

Guopeng Teng, Gongde Lin, Pengfan Wei, Lizhi Li, Hongyuan Chen, Qingquan Chen and Qiuyuan Lin

{"title":"FEN1-assisted LAMP for specific and multiplex detection of pathogens associated with community-acquired pneumonia†","authors":"Guopeng Teng, Gongde Lin, Pengfan Wei, Lizhi Li, Hongyuan Chen, Qingquan Chen and Qiuyuan Lin","doi":"10.1039/D4AN01516A","DOIUrl":"10.1039/D4AN01516A","url":null,"abstract":"Lower respiratory tract infections (LRITs), including community-acquired pneumonia (CAP), are the fifth leading cause of death worldwide over the last ten years, posing a serious threat to global healthcare. Conventional laboratory assays for detecting pathogens are hindered by complicated procedures, a long turnaround time and a lack of multiplex detection capabilities. In this study, a flap-endonuclease 1 (FEN1)-assisted loop-mediated isothermal amplification (LAMP) method was designed, and an assay based on this method was developed to identify three leading pathogens for CAP, namely, Streptococcus pneumoniae, Mycoplasma pneumoniae and Haemophilus influenzae. FEN1-assisted LAMP utilized a sequence-specific probe with a flap structure to generate an amplified signal, demonstrating high specificity and sensitivity with a low limit of detection (100 copies per μL). Based on the cleavage of flap probes by FEN1, our assay was able to detect three pathogens in a single reaction. This method is highly consistent with the polymerase chain reaction (PCR) in clinical sample testing. This simple, specific and multiple detection method has the potential to identify CAP and could be applied to detect other pathogen infections.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1419-1426"},"PeriodicalIF":3.6,"publicationDate":"2025-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/an/d4an01516a?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143518287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Raman spectroscopy of ovarian and peritoneal tissue in the assessment of ovarian cancer†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-27 DOI: 10.1039/D4AN01293C

Diana Frimpong, Angela C. Shore, Benjamin Gardner, Claire Newton, Joya Pawade, Jonathan Frost, Laura Atherton and Nick Stone

{"title":"Raman spectroscopy of ovarian and peritoneal tissue in the assessment of ovarian cancer†","authors":"Diana Frimpong, Angela C. Shore, Benjamin Gardner, Claire Newton, Joya Pawade, Jonathan Frost, Laura Atherton and Nick Stone","doi":"10.1039/D4AN01293C","DOIUrl":"10.1039/D4AN01293C","url":null,"abstract":"During post chemotherapy surgery for ovarian cancer, it is important to ensure that any residual disease is carefully assessed and removed. The assessment remains subjective, despite clear evidence of the benefits of complete macroscopic resection. In this work, we have considered Raman spectroscopy as a possible tool for residual disease assessment by exploring its ability to correctly classify ovarian cancer from benign and borderline tissues. Samples from seventy-three participants were analysed (n = 20 benign, n = 11 borderline and n = 42 cancer) using a multivariate analysis model. All models shown utilised validation with leave one participant out cross-validation. In ovarian tissue this model achieved 94% sensitivity and 98% specificity for prediction of cancer from benign and 98% sensitivity and 89% specificity for prediction of cancer from borderline. Thorough assessment of the surrounding peritoneal tissues is extremely important. For these peritoneal tissues taken from participants with advanced ovarian cancer, the model achieved 78% sensitivity and 84% specificity for prediction of cancerous peritoneum from benign peritoneum in participants who had primary surgery and 68% sensitivity and 81% specificity in participants who had post chemotherapy surgery. This demonstrates viability of Raman spectroscopy for assessment of ovarian cancer.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1303-1309"},"PeriodicalIF":3.6,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.rsc.org/en/content/articlepdf/2025/an/d4an01293c?page=search","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Exploring the potential of Fourier transform-infrared spectroscopy of urine for non-invasive monitoring of inflammation associated with a kidney transplant†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-27 DOI: 10.1039/D4AN01459F

Elie Sarkees, Vincent Vuiblet, Fayek Taha and Olivier Piot

{"title":"Exploring the potential of Fourier transform-infrared spectroscopy of urine for non-invasive monitoring of inflammation associated with a kidney transplant†","authors":"Elie Sarkees, Vincent Vuiblet, Fayek Taha and Olivier Piot","doi":"10.1039/D4AN01459F","DOIUrl":"10.1039/D4AN01459F","url":null,"abstract":"The global rise of end-stage renal disease is leading to an increase in kidney transplants. Graft survival is dependent on the occurrence of inflammation which can lead to cases of rejection. Traditional laboratory analyses often lack accuracy, and graft biopsies – the current gold standard – are considered invasive and risky. This highlights an unmet need for innovative diagnostic and monitoring methods of graft rejection and inflammation. This study explores the potential of Fourier-transform infrared spectroscopy of fresh urine for diagnosing kidney transplant inflammation. Urine samples were collected from kidney transplant patients who were under regular surveillance. An unsupervised method of spectral data analysis, especially Uniform Manifold Approximation and Projection (UMAP), was initially employed. However, it was unable to reveal a clear distinction between control and pathological conditions. Subsequently, two machine learning models – SVM and gradient boosting – were employed to categorise participants into pathologic or control groups, achieving a diagnostic accuracy of 77.78%. This study also evaluated other factors that could affect model performance, including urine biochemical composition, type of inflammation, and patient's medication history. The inherent variability of urine, attributed to factors such as diet and medications, poses challenges to identifying robust spectroscopic markers. Nevertheless, mid-infrared spectroscopy offers a promising, non-invasive approach for diagnosing kidney transplant disorders. Further research is essential to provide more advanced prediction models and meet the criteria for potential clinical deployment.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1427-1435"},"PeriodicalIF":3.6,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143518288","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 CRISPR/Cas13a-based and hybridization chain reaction coupled evanescent wave biosensor for SARS-CoV-2 gene detection†

IF 3.6 3区化学

Analyst Pub Date : 2025-02-27 DOI: 10.1039/D4AN01584C

Yang Li, Yikan Zhao, Zhihao Yi and Shitong Han

{"title":"A CRISPR/Cas13a-based and hybridization chain reaction coupled evanescent wave biosensor for SARS-CoV-2 gene detection†","authors":"Yang Li, Yikan Zhao, Zhihao Yi and Shitong Han","doi":"10.1039/D4AN01584C","DOIUrl":"10.1039/D4AN01584C","url":null,"abstract":"Timely and accurate diagnosis of RNA viruses, represented by the SARS-CoV-2, is the foundation for protecting people from health threats. Currently, direct detection of viral RNA genes remains the most accurate method. Herein, a rapid, ultrasensitive, and no heating equipment required CRISPR/Cas13a based evanescent wave fluorescence biosensing platform for quantitative detection of the SARS-CoV-2 gene is reported. The collateral effect of CRISPR/Cas13a for RNA is combined with a self-driven enzyme-free hybridization chain reaction (HCR) as a signal amplification step. When the initiator RNA strand is cleaved by Cas13a, the downstream signal amplification induced by HCR is blocked, and a multiple crRNA strategy is used to enhance the cleavage efficiency. The newly designed HCR assemblies are captured by the cDNA-modified optical fiber and generate a higher-intensity fluorescence signal induced by the evanescent field. The CRISPR/Cas13a-HCR evanescent wave fluorescence biosensing platform is capable of detection of SARS-CoV-2 with a LOD of 0.47 copies per μL and the detection time is within 35 min. The spike recovery tests in saliva and high specificity have demonstrated the potential of this method for point-of-care diagnosis. This method is also suitable for the detection of other RNA viruses, without the need to alter any design of the HCR component, and only the corresponding crRNA needs to be replaced.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 7","pages":" 1367-1376"},"PeriodicalIF":3.6,"publicationDate":"2025-02-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143506827","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