Talanta最新文献

{"title":"A novel electrochemical approach for sensitive and simultaneous detection of 7,8-dihydro-8-oxoadenine and 2-hydroxyadenine based on a screen-printed carbon electrode modified with carbon-encapsulated Fe3O4","authors":"Yue Wang ,&nbsp;Fei Yu ,&nbsp;Qing-Hua Liu ,&nbsp;Cai-Yun Wang ,&nbsp;Guo-Yuan Zhu ,&nbsp;Li-Ping Bai ,&nbsp;Ke-Ying Guo ,&nbsp;Zhi-Hong Jiang ,&nbsp;Wei Zhang","doi":"10.1016/j.talanta.2025.128584","DOIUrl":"10.1016/j.talanta.2025.128584","url":null,"abstract":"<div><div>Excessive reactive oxygen species attack DNA, resulting in the formation of oxidative guanine (oxoG) and oxidative adenine (oxoA) as significant types of oxidative DNA damage. Despite the similar carcinogenic potential and content levels of oxoA, there is still a lack of comprehensive studies and detection methodologies compared to oxoG. Herein, a novel electrochemical approach for sensitive and simultaneous detection of 7,8-dihydro-8-oxoadenine (8-oxoA) and 2-hydroxyadenine (2-oxoA) was first reported and designed using screen-printed carbon electrode (SPCE) modified with carbon-encapsulated Fe₃O₄ (Fe₃O₄@C). The SPCE modified with Fe₃O₄@C (Fe₃O₄@C/SPCE) exhibited excellent electrocatalytic performance for the simultaneous detection of 8-oxoA and 2-oxoA without requiring any enzymes. Although 8-oxoA and 2-oxoA are a pair of isomers, their oxidation peaks could be separated using differential pulse voltammetry. Under the optimal conditions, the corresponding limits of detection were 45.3 nM and 34.5 nM for 8-oxoA and 2-oxoA (S/N = 3), demonstrating high sensitivity and a wide linear range from 0.05 μM to 100 μM. The proposed electrochemical sensor was also applied to oxoA analysis in rat serum samples with satisfactory recovery values. Overall, Fe₃O₄@C/SPCE displayed a promising oxoA assay platform, which expands the choice of biomarkers in purine oxidation and contributes to describing DNA oxidative damage more comprehensively and accurately.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128584"},"PeriodicalIF":5.6,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancement of doxorubicin monitoring with a DNA fragmentation strategy for SYBR Green I-based aptamer biosensors","authors":"Luke Wei,&nbsp;Yuxin Hu,&nbsp;Jiamin Wu,&nbsp;Jing Mao,&nbsp;Tianqin Yin,&nbsp;Jieqiong Qiu","doi":"10.1016/j.talanta.2025.128601","DOIUrl":"10.1016/j.talanta.2025.128601","url":null,"abstract":"<div><div>Doxorubicin (DOX), a widely utilized chemotherapeutic antibiotic, increasingly threatens environmental and public health due to its persistence in marine ecosystems, bioaccumulation in organisms, and contamination of agricultural systems. In response to the urgent demand for effective DOX detection, we have developed a label-free fluorescent aptamer biosensor (FAB) that employs SYBR Green I (SG I)-mediated signal amplification. A key innovation in this design is the strategic fragmentation of the 41-mer complementary strand (CP) of the DOX aptamer into three 11-mer fragments. It significantly mitigates nonspecific SG I binding interference caused by the longer ssDNA CP, enhancing the fluorescence signal ratio (F_dsDNA/F_ssDNA) from 4.6-fold to 12.6-fold. Upon DOX introduction, competitive binding between DOX, SG I, and DNA further suppresses background noise, yielding an exceptional fluorescence quenching ratio (F₀/F) of 52.8. The optimized FAB demonstrates ultra-sensitive DOX detection, with a detection limit of 1.41 nM and a linear response between 1.41 nM and 300 nM. The recoveries of DOX spiked into pond water and drinking water samples ranged from 95.1 % to 101.8 %, with relative standard deviations (RSDs) below 3.46 %, demonstrating negligible matrix effects and confirming the FAB method's robustness for environmental monitoring applications. This strategy synergistically enhances signal transduction efficiency, and strengthens detection reliability, all while maintaining operational simplicity. As a result, this FAB for DOX detection holds transformative potential, poised to revolutionize biosensor technology in the fields of food safety, environmental monitoring, and biosecurity.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128601"},"PeriodicalIF":5.6,"publicationDate":"2025-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144686774","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-targeted metabolomics and machine learning reveal metabolic dysregulation in lymph node tuberculosis","authors":"Peijun Chen ,&nbsp;Yuehui Yu ,&nbsp;Ying Zhang ,&nbsp;Gaoyi Yang","doi":"10.1016/j.talanta.2025.128583","DOIUrl":"10.1016/j.talanta.2025.128583","url":null,"abstract":"<div><h3>Background</h3><div>Lymph node tuberculosis (LNTB) is the most prevalent form of extrapulmonary tuberculosis; however, differentiating it from non-LNTB remains challenging due to overlapping clinical features and suboptimal diagnostic methods. Current diagnostic methods for LNTB lack both sensitivity and specificity. This study aimed to characterize the metabolic differences between LNTB and non-LNTB patients, elucidate the pathological mechanisms underlying LNTB, and identify diagnostic biomarkers using machine learning models.</div></div><div><h3>Methods</h3><div>Serum samples from 40 LNTB patients and 30 non-LNTB patients were analyzed using ultra-high-performance liquid chromatography-mass spectrometry. Differential metabolites were identified based on a variable importance in projection &gt;1, false discovery rate-adjusted p-value &lt;0.05. Pathway enrichment analysis was performed using the Kyoto Encyclopedia of Genes and Genomes (KEGG). Machine learning, including support vector machines and random forest, were employed to screen for diagnostic biomarkers, which were validated by receiver operating characteristic curves.</div></div><div><h3>Results</h3><div>Among the 1294 detected metabolites, 89 exhibited significant differences between the two groups. By integrating KEGG enrichment with topological analysis, phenylalanine, tyrosine, and tryptophan biosynthesis possessed the highest impact, followed by phenylalanine metabolism, and aminoacyl-tRNA biosynthesis. Machine learning identified four biomarkers: Leu-Ala [area under the curve (AUC) = 0.8292], evodiamine (AUC = 0.7558), fenazaquin (AUC = 0.7175), and acetol (AUC = 0.7117). Leu-Ala demonstrated the highest diagnostic accuracy, with a sensitivity of 73.5 % and specificity 86.7 % at a cutoff value of 0.62.</div></div><div><h3>Conclusions</h3><div>Untargeted metabolomics revealed dysregulation in the biosynthesis of phenylalanine, tyrosine, and tryptophan, phenylalanine metabolism, as well as in aminoacyl-tRNA biosynthesis in LNTB. Additional, Leu-Ala was identified as a novel diagnostic biomarker. The integrating of metabolomics with machine learning presents a promising approach for LNTB detection, though larger validation studies are necessary.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128583"},"PeriodicalIF":5.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672592","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization and comparison of amino group derivatization reagents for sensitive and isomer-selective LC-MS/MS analysis of tyrosine-derived biomarkers of oxidative and nitrosative stress","authors":"Yusuke Iwasaki ,&nbsp;Hironori Saito ,&nbsp;Kangrong Chen ,&nbsp;Hu Leqi ,&nbsp;Natsuki Koshiishi ,&nbsp;Keisuke Mochizuki ,&nbsp;Rie Ito ,&nbsp;Hiroshi Akiyama","doi":"10.1016/j.talanta.2025.128596","DOIUrl":"10.1016/j.talanta.2025.128596","url":null,"abstract":"<div><div>Reactive oxygen species and reactive nitrogen species play roles in the pathogenesis of numerous diseases, but their presence is often assessed indirectly via analysis of specific biomarkers. Tyrosine-derived compounds such as ortho-tyrosine, meta-tyrosine, 3-chlorotyrosine, and 3-nitrotyrosine are considered markers of oxidative and nitrosative stress. However, these biomarkers are typically present in extremely low concentrations (in the low nanomolar range) and have structural isomers, making accurate quantification challenging. The aim of this study was to improve the sensitivity and chromatographic separation of these analytes using various commercially available derivatization reagents targeting amino groups. Selected reaction monitoring transitions were optimized by flow injection analysis of derivatized standards, and structural isomers were evaluated using LC-MS/MS. A Box-Behnken design and response surface methodology were employed to determine the optimal derivatization conditions for each reagent. Among the reagents tested, diethyl ethoxymethylenemalonate was most effective, enabling complete isomer separation and approximately 1000-fold signal enhancement compared with non-derivatized analytes, with a limit of quantification reaching 5 nM. The results of this study highlight the importance of selecting an appropriate derivatization strategy for sensitive and selective quantification of tyrosine-based biomarkers and offer a promising approach for the accurate assessment of oxidative and nitrosative stress in biological samples.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128596"},"PeriodicalIF":5.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144694767","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pine wood species identification based on random forest transformer and near infrared spectroscopy","authors":"Chengwu Chen,&nbsp;Yuan da Qi,&nbsp;Zijian Qin,&nbsp;Yiwei Li,&nbsp;Yaoxiang Li","doi":"10.1016/j.talanta.2025.128599","DOIUrl":"10.1016/j.talanta.2025.128599","url":null,"abstract":"<div><div>In the wood trade market, inferior timber is frequently mislabeled as a superior timber for higher profits. To effectively avoid this and circumvent unnecessary economical losses, this study proposes a pine species identification based on Random Forest Transformer and near infrared spectroscopy to identify the three kinds of pine wood, namely, <em>Pinus koraiensis</em> (red pine), <em>Pinus sylvestris</em> (scotch pine) and <em>Larix gmelinii</em> (larch). Preprocessing methods were applied to the spectra data of the pine wood samples. Feature extraction of the near infrared spectroscopy was done with Random Forest algorithm. Then the identification models of pine woods were developed based on the Random Forest Transformer, Linear Discriminant Analysis (LDA), Support Vector Machines (SVM) and Convolution Neural Networks (CNN). Results showed that the method proposed for pine species identification could quickly and effectively determine the category of pine wood with discrimination accuracy of 98.39 %, which was improved by 3.23 %, 1.62 % and 1.62 % compared with the results of LDA, SVM, and CNN, respectively. Furthermore, the proposed method holds considerable scientific significance and reference value for non-destructive testing applications, not only in the identification of other wood species but also in fields such as traditional Chinese medicine, food science, and agriculture.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128599"},"PeriodicalIF":5.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144686775","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Agarose-loaded mesoporous silica nanocomposite embedded in a polyether sulfone substrate for thin film solid-phase microextraction: An approach for narcotic drugs extraction from eye tears in a multi-channel microfluidic device prior to liquid chromatography-tandem mass spectrometry","authors":"Shirin Yadegari ,&nbsp;Shahram Seidi ,&nbsp;Reyhaneh Namdari","doi":"10.1016/j.talanta.2025.128592","DOIUrl":"10.1016/j.talanta.2025.128592","url":null,"abstract":"<div><div>Herein, a solid-phase microextraction method was developed based on an agarose-loaded mesoporous silica nanocomposite embedded in a polyether sulfone thin film, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The microextraction was performed using a multi-channel microfluidic device incorporating the prepared nanocomposite thin film to extract morphine (MOR), heroin (HER), codeine (COD), and methamphetamine (MET) from eye tear samples. Field emission scanning electron microscopy (FE-SEM), Fourier-transform infrared spectroscopy (FT-IR), Brunauer–Emmett–Teller (BET), thermal gravimetric analysis (TGA), and transmission electron microscopy (TEM) were employed to characterize the nanocomposite. The influence of various parameters on the extraction efficiency—including sample pH, sample flow rate, nanoparticle and polyether sulfone amounts, eluent type and volume, and eluent flow rate—was systematically studied. Under optimized conditions, the limits of detection (LOD) were 0.07 and 0.06 ng mL⁻¹ for COD and HER, respectively, and 0.1 ng mL⁻¹ for MET and MOR. The matrix effect, ranging from 8.85 % to 45.64 %, was evaluated using box-whisker plots. Intra-day and inter-day RSDs% were below 4.5 % and 4.0 %, respectively. Calibration curves showed linearity over 0.5–80.0 ng mL⁻¹ for MET, 0.2–80.0 ng mL⁻¹ for COD, and 0.4–80.0 ng mL⁻¹ for MOR and HER. Relative recovery (RR%) values assessing accuracy in tear samples ranged from 90.0 % to 112.0 %. Greenness and Blueness scores of 0.72 and 57.5, respectively, demonstrate that the suggested approach is both sustainable and applicable. These findings indicate that this method is a suitable alternative for measuring trace amounts of narcotic drugs in tear samples.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128592"},"PeriodicalIF":5.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Smart and sustainable 3D-printed electrochemical device for diclofenac remediation and monitoring in water","authors":"Gabriella Iula ,&nbsp;Ada Raucci ,&nbsp;Lorenzo Antonelli ,&nbsp;Panagiota M. Kalligosfyri ,&nbsp;Massimo Giuseppe De Cesaris ,&nbsp;Nina Felli ,&nbsp;Concetta Di Natale ,&nbsp;Stefano Cinti ,&nbsp;Alessandra Gentili","doi":"10.1016/j.talanta.2025.128597","DOIUrl":"10.1016/j.talanta.2025.128597","url":null,"abstract":"<div><div>In recent years, emerging contaminants in water, such as pesticides and pharmaceuticals, have gained significant attention, underscoring the need for innovative purification and monitoring solutions. Diclofenac, a widely used nonsteroidal anti-inflammatory drug, has become an important environmental contaminant due to its widespread use and the poor efficiency of conventional wastewater treatment systems. This study focuses on the development of an innovative all-in-one 3D printed device for small-scale monitoring and remediation of emerging contaminants such as diclofenac, with the potential to be adapted to others as well. The main novelty of this work lies in the design of a compact and portable platform that not only enables the detection of diclofenac in real time but also assesses the quality of its remediation. The 3D-printed device integrates a flexible, screen-printed sensor on a polyester substrate for diclofenac detection, along with recycled cellulose acetate particles functionalized with activated carbon (20 % w/w) as the remediation material. Combining electrochemical sensing technologies with a lightweight and cost-effective three-dimensional configuration, the platform offers a highly efficient and easy-to-use solution for monitoring and optimizing remediation processes. The device provides real-time feedback on diclofenac concentrations, evaluating remediation efficiency under practical, real-world conditions. Results showed that the all-in-one platform achieved a detection limit for diclofenac of 0.1 μM and demonstrated a remediation efficiency of about 53 %. This innovative 3D system represents a significant advance in the industry, offering a versatile and scalable solution to improve the management of pharmaceutical contaminants in water resources, with broader applications for various emerging pollutants.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128597"},"PeriodicalIF":5.6,"publicationDate":"2025-07-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144653455","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of complementary enrichment strategies for analysis of N-linked intact glycopeptides and potential site-specific glycoforms in Alzheimer's disease","authors":"Zhenpeng Deng ,&nbsp;Yan Wang ,&nbsp;Xiaoyan Liu ,&nbsp;Xingfa Ren ,&nbsp;Weibing Zhang ,&nbsp;Mingming Dong ,&nbsp;Yusong Ge ,&nbsp;Yongliang Yu ,&nbsp;Mingliang Ye","doi":"10.1016/j.talanta.2025.128595","DOIUrl":"10.1016/j.talanta.2025.128595","url":null,"abstract":"<div><div>Protein glycosylation is a critical post-translational modification, and knowledge of site-specific glycoforms is essential for developing biomarkers and therapeutic drugs. Although LC-MS/MS-based glycoproteomics strategies enable the identification of site-specific glycoforms at proteomics scale, their coverage is still low largely because of the poor glycopeptide enrichment performance. HILIC is thought to allow \"unbiased\" enrichment of intact glycopeptides, and it is broadly used to analyze the site-specific glycoforms at proteomics scale. To maximize glycopeptide capturing, the samples are always loaded onto HILIC with high acetonitrile (ACN) content (typically 80 %). In this study, we found that some HILIC columns could effectively capture glycopeptides at around 70 % ACN. We further demonstrated that the system could identify the highly hydrophilic glycopeptides that can not be identified by conventional methods. The excellent complementarity of 70 % ACN enrichment methods greatly enhances the coverage (&gt;20 %) of N-glycoproteome identification in human blood. The developed methods were further applied to investigate the N-glycosylation changes in the plasma of Alzheimer's disease (AD) and mild cognitive impairment (MCI) patients. It was found that fucosylated glycopeptides were up-regulated and sialylated glycopeptides were down-regulated as the disease progressed. Altered glycosylation patterns were detected for a number of site-specific glycoforms, which serve as potentially interesting targets for further glycosylation-based AD progression. Our results reveal that using complementary strategies offers a comprehensive approach to studying N-glycoproteomics, paving the way for in-depth glycoproteomics analysis.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128595"},"PeriodicalIF":5.6,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144653456","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Nanozyme-mediated smartphone platform for colorimetric detection of total leucomalachite green and malachite green","authors":"Xinru Nan ,&nbsp;Xiaoqing Yang ,&nbsp;Qiaowen Lei ,&nbsp;Yunxiao Zhu ,&nbsp;Chenghao Liu ,&nbsp;Jing Liao ,&nbsp;Chendi Qin ,&nbsp;Anqi Liu ,&nbsp;Man Zhou ,&nbsp;Xudong Wang ,&nbsp;Qitong Huang ,&nbsp;Xiaoqing Yi","doi":"10.1016/j.talanta.2025.128594","DOIUrl":"10.1016/j.talanta.2025.128594","url":null,"abstract":"<div><div>As a parasiticide, malachite green (MG) and its primary metabolite, leucomalachite green (LMG), are widely used in aquaculture to boost fishery yields. However, these compounds pose significant risks to both the food chain and human health. This study presents an innovative colorimetric sensing platform utilizing MnO₂ nanozymes for rapid and precise quantification of total LMG and MG in aquatic products. The innovation of platform lies in three key aspects: (1) development of an environmentally benign synthesis protocol for high-performance MnO₂ nanozymes using gelatin as a biological template and KMnO₄ as precursor; (2) exploitation of the distinctive oxidative catalytic properties of MnO₂ nanozymes, enabling in situ conversion of LMG to MG within 1 min under substrate-free conditions, coupled with the characteristic absorption peak of MG at 618 nm for highly sensitive and selective detection (LOD: 33.00 μg/kg); and (3) seamless integration with smartphone technology to create a cost-effective, portable, and user-friendly intelligent detection system (LOD: 120.08 μg/kg). The actual sample analysis results show that the integrated detection platform exhibits excellent analytical performance in complex matrices such as fish muscle, shrimp tissue, and fish pond water. The spike recovery rate of UV–vis spectroscopy and smartphone colorimetric analysis reaches 97.92 %–106.50 % and 97.21 %–104.17 %, respectively. It has high detection efficiency and real-time monitoring capabilities, providing a powerful solution for aquatic product safety assessment, public health protection and environmental monitoring.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128594"},"PeriodicalIF":5.6,"publicationDate":"2025-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144672589","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid and simple lateral flow detection for ADH1B and ALDH2 genotyping: PCR-restriction digestion-adapter ligation-lateral flow (PRALL) assay","authors":"Naoto Shirasu,&nbsp;Shin'ichiro Yasunaga","doi":"10.1016/j.talanta.2025.128588","DOIUrl":"10.1016/j.talanta.2025.128588","url":null,"abstract":"<div><div>Single nucleotide polymorphisms (SNPs) in <em>alcohol dehydrogenase 1B</em> (<em>ADH1B</em>) and <em>aldehyde dehydrogenase 2</em> (<em>ALDH2</em>) significantly impact ethanol metabolism and alcohol-related health risks. Rapid and simple genotyping methods for these genes are crucial for clinical diagnostics and point-of-care testing (POCT), but existing techniques often require complex procedures or specialized equipment. We developed and validated a novel genotyping method, termed a PCR-restriction digestion-adapter ligation-lateral flow (PRALL) assay. The PRALL assay integrates: (i) template preparation from buccal swab, (ii) duplex PCR amplification using biotinylated primers incorporating a <em>Bsl</em>I recognition site, (iii) simultaneous <em>Bsl</em>I restriction digestion and allele-specific ligation of hapten-labeled adapters in a single tube, and (iv) visual detection by lateral flow assay (LFA). The method was successfully validated using model templates and human genomic DNA. The complete PRALL assay, from crude lysate preparation to visual result, was performed in approximately 1 h. The assay demonstrated high sensitivity (limit of detection: ∼100 pg genomic DNA, corresponding to ∼15 allele copies) and achieved 100 % concordance with Sanger genotyping for both SNPs across 25 human samples. The unique integration of streamlined enzymatic steps with an instrument-free visual LFA readout overcomes key limitations of previous methods. This makes PRALL a promising and practical tool for facilitating personalized alcohol risk assessment and enhancing clinical diagnostics, demonstrating strong potential for deployment in various settings, including those closer to the point of care.</div></div>","PeriodicalId":435,"journal":{"name":"Talanta","volume":"297 ","pages":"Article 128588"},"PeriodicalIF":5.6,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144634623","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}