Pub Date : 2026-02-22Epub Date: 2026-01-15DOI: 10.1016/j.aca.2026.345100
Daria Tikhonova , Evgeny Andreev , Assel Akhmetova , Nadezda Meshcheryakova , Olga Zaborova , Liliya Mukhametova , Rugiya Aliyeva , Elena Kravchenko , Elizaveta Boravleva , Sergei Dubkov , Sergei Eremin , Igor Yaminsky , Alexander Nechaev , Elena Zavyalova
Rapid specific sensors, ready for several particles’ detection per probe, are essential for disease diagnostics at the early stages before the symptom manifestation. Infectious diseases represent a prominent example where the early diagnosis is necessary both for a proper treatment of the patient as well as preventing the infection spreading. Here, a composite track-etched membrane with a 3D surface was used to create a biosensor specific to influenza A virus. The membrane allows concentration of the analyte on the surface, whereas the spike-like 3D surface covered with Ag layer serves as a substrate for surface-enhanced Raman spectroscopy (SERS) enhancing Raman scattering by 106-fold. The SERS-active coating is stable even in biological medium, being resistant to salts and proteins. A DNA aptamer with broad specificity to various influenza A strains was used as a recognition element, which was attached to the metal surface. As a result, the sensor can determine both human and avian influenza A viruses, covering all risky viral strains. The limit of detection (LoD) was as low as 120–2000 viral particles in mL. The LoDs are comparable with the LoDs of polymerase chain reaction with reverse transcription when the time of analysis is about 10 min. The biosensor allows rapid, specific and accurate detection of influenza A viruses in the nasal swabs.
{"title":"Specific determination of influenza virus A by SERS-active spike-like track-etched membranes","authors":"Daria Tikhonova , Evgeny Andreev , Assel Akhmetova , Nadezda Meshcheryakova , Olga Zaborova , Liliya Mukhametova , Rugiya Aliyeva , Elena Kravchenko , Elizaveta Boravleva , Sergei Dubkov , Sergei Eremin , Igor Yaminsky , Alexander Nechaev , Elena Zavyalova","doi":"10.1016/j.aca.2026.345100","DOIUrl":"10.1016/j.aca.2026.345100","url":null,"abstract":"<div><div>Rapid specific sensors, ready for several particles’ detection per probe, are essential for disease diagnostics at the early stages before the symptom manifestation. Infectious diseases represent a prominent example where the early diagnosis is necessary both for a proper treatment of the patient as well as preventing the infection spreading. Here, a composite track-etched membrane with a 3D surface was used to create a biosensor specific to influenza A virus. The membrane allows concentration of the analyte on the surface, whereas the spike-like 3D surface covered with Ag layer serves as a substrate for surface-enhanced Raman spectroscopy (SERS) enhancing Raman scattering by 10<sup>6</sup>-fold. The SERS-active coating is stable even in biological medium, being resistant to salts and proteins. A DNA aptamer with broad specificity to various influenza A strains was used as a recognition element, which was attached to the metal surface. As a result, the sensor can determine both human and avian influenza A viruses, covering all risky viral strains. The limit of detection (LoD) was as low as 120–2000 viral particles in mL. The LoDs are comparable with the LoDs of polymerase chain reaction with reverse transcription when the time of analysis is about 10 min. The biosensor allows rapid, specific and accurate detection of influenza A viruses in the nasal swabs.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345100"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145993297","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-22Epub Date: 2026-01-16DOI: 10.1016/j.aca.2026.345112
Yang Li , Yu He , Weijie Zhang , Mengqing Yang , Wei Yu , Zhenbin Zhang
Background
Protein N-terminal acetylation (Nt-acetylation) is a widespread and essential modification in both eukaryotes and prokaryotes. To achieve in-depth profiling of Nt-acetylome, enrichment of Nt-acetylated peptides prior to mass spectrometry (MS) analysis is crucial, as their inherently low ionization efficiency is further suppressed in complex peptide mixtures. However, current methods are constrained by low enrichment selectivity and insufficient proteolytic digestion efficiency toward Nt-acetylated peptides.
Results
Herein, we present a novel and effective enrichment method involving the specific blocking of lysine ε-amino groups by guanidination and long alkyl chain tagging to N-terminal/internal peptides with free α-amino groups. Due to their enhanced hydrophobicity, the alkylated N-terminal/internal peptides could be efficiently depleted via a C18 column, thus achieving the negative enrichment of Nt-acetylated peptides. Specific guanidination of lysine residues ensures high enrichment selectivity and efficient tryptic digestion of Nt-acetylated peptides. In a single-shot analysis of HeLa cells, our method yielded a significant 4-fold increase in the identification number of Nt-acetylated peptides compared to direct analysis. When coupled with high-pH C18 fractionation, this approach identified 4957 unique Nt-acetylated peptides, corresponding to 3042 acetylated N-termini and 902 putatively neo-acetylated N-termini, markedly expanding the identification coverage of current Nt-acetylation dataset. Furthermore, this method was successfully applied to the quantification of N-terminal acetylome in hippocampal tissues from Alzheimer's disease (AD) mice modeled by amyloid-beta (Aβ) hippocampal injection.
Significance
Our method exhibits high enrichment selectivity, efficient proteolytic efficiency and minimal bias toward Nt-acetylated peptides in complex samples. This robust and versatile strategy offers an efficient alternative for comprehensive profiling of Nt-acetylome, thereby facilitating deeper insights into the physiological and pathological functions of Nt-acetylation across diverse biological systems.
{"title":"Specific lysine guanidination and long alkyl chain tagging-assisted negative enrichment strategy enables comprehensive profiling of protein N-terminal acetylome","authors":"Yang Li , Yu He , Weijie Zhang , Mengqing Yang , Wei Yu , Zhenbin Zhang","doi":"10.1016/j.aca.2026.345112","DOIUrl":"10.1016/j.aca.2026.345112","url":null,"abstract":"<div><h3>Background</h3><div>Protein N-terminal acetylation (Nt-acetylation) is a widespread and essential modification in both eukaryotes and prokaryotes. To achieve in-depth profiling of Nt-acetylome, enrichment of Nt-acetylated peptides prior to mass spectrometry (MS) analysis is crucial, as their inherently low ionization efficiency is further suppressed in complex peptide mixtures. However, current methods are constrained by low enrichment selectivity and insufficient proteolytic digestion efficiency toward Nt-acetylated peptides.</div></div><div><h3>Results</h3><div>Herein, we present a novel and effective enrichment method involving the specific blocking of lysine ε-amino groups by guanidination and long alkyl chain tagging to N-terminal/internal peptides with free α-amino groups. Due to their enhanced hydrophobicity, the alkylated N-terminal/internal peptides could be efficiently depleted via a C18 column, thus achieving the negative enrichment of Nt-acetylated peptides. Specific guanidination of lysine residues ensures high enrichment selectivity and efficient tryptic digestion of Nt-acetylated peptides. In a single-shot analysis of HeLa cells, our method yielded a significant 4-fold increase in the identification number of Nt-acetylated peptides compared to direct analysis. When coupled with high-pH C18 fractionation, this approach identified 4957 unique Nt-acetylated peptides, corresponding to 3042 acetylated N-termini and 902 putatively neo-acetylated N-termini, markedly expanding the identification coverage of current Nt-acetylation dataset. Furthermore, this method was successfully applied to the quantification of N-terminal acetylome in hippocampal tissues from Alzheimer's disease (AD) mice modeled by amyloid-beta (Aβ) hippocampal injection.</div></div><div><h3>Significance</h3><div>Our method exhibits high enrichment selectivity, efficient proteolytic efficiency and minimal bias toward Nt-acetylated peptides in complex samples. This robust and versatile strategy offers an efficient alternative for comprehensive profiling of Nt-acetylome, thereby facilitating deeper insights into the physiological and pathological functions of Nt-acetylation across diverse biological systems.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345112"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145972580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-22Epub Date: 2026-01-12DOI: 10.1016/j.aca.2026.345096
Caiyuan Liao , Fayu Wang , Long Fang , Rong Zou , Yaping Wang , Gang Chang , Hanping He
A field-effect transistor based on donor-acceptor (D-A) copolymer/graphene (GR) blends was successfully developed, with systematic optimizations implemented for key parameters including the concentration of polymeric monomers, selection of blending materials, mass ratio of blended components, and post-deposition annealing temperature of the composite films. The D-A copolymer was synthesized by Stille polycondensation with isoindigo as the acceptor (A) unit and thieno [3,2-b] thiophene as the donor (D) unit. The PIID-C10-TT/GR blend with a ratio of 95/5 exhibited the optimal performance in the device, achieving a carrier mobility of 0.123 ± 0.014 cm2/(V·s), a subthreshold swing of 308.00 ± 34.87 mV/dec, and an on/off ratio of 1.27 × 103. The novel Ab/ADP/GR-FET biosensor exhibited excellent sensing performance for the detection of the EpCAM biomarker, with a detection range of 0.05–20 ng/mL, a lower detection limit (LOD) of 24 pg/mL, and high selectivity. Notably, it demonstrated robust detection capability for circulating tumor cells (MCF-7), showing a linear response range from 5 to 50 cells. These studies indicated that the electrochemical field-effect transistor (EFET) based on the PIID-C10-TT/GR blend as a semiconductor material holds significant potential for biosensing applications.
{"title":"Preparation and application of the transistor based on D-A isoindigo copolymer/GR blend for the detection of circular tumor cell","authors":"Caiyuan Liao , Fayu Wang , Long Fang , Rong Zou , Yaping Wang , Gang Chang , Hanping He","doi":"10.1016/j.aca.2026.345096","DOIUrl":"10.1016/j.aca.2026.345096","url":null,"abstract":"<div><div>A field-effect transistor based on donor-acceptor (D-A) copolymer/graphene (GR) blends was successfully developed, with systematic optimizations implemented for key parameters including the concentration of polymeric monomers, selection of blending materials, mass ratio of blended components, and post-deposition annealing temperature of the composite films. The D-A copolymer was synthesized by Stille polycondensation with isoindigo as the acceptor (A) unit and thieno [3,2-b] thiophene as the donor (D) unit. The PIID-C10-TT/GR blend with a ratio of 95/5 exhibited the optimal performance in the device, achieving a carrier mobility of 0.123 ± 0.014 cm<sup>2</sup>/(V·s), a subthreshold swing of 308.00 ± 34.87 mV/dec, and an on/off ratio of 1.27 × 10<sup>3</sup>. The novel Ab/ADP/GR-FET biosensor exhibited excellent sensing performance for the detection of the EpCAM biomarker, with a detection range of 0.05–20 ng/mL, a lower detection limit (LOD) of 24 pg/mL, and high selectivity. Notably, it demonstrated robust detection capability for circulating tumor cells (MCF-7), showing a linear response range from 5 to 50 cells. These studies indicated that the electrochemical field-effect transistor (EFET) based on the PIID-C10-TT/GR blend as a semiconductor material holds significant potential for biosensing applications.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345096"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145957346","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-22Epub Date: 2026-01-14DOI: 10.1016/j.aca.2026.345103
Yicheng Yao , Liudi Ji , Peng Hu , Xiaoming Zhu , Junxing Hao , Kangbing Wu
Background
Carbendazim (CBZ), is a benzimidazole fungicide and widely used for controlling fruit and vegetable diseases, which poses significant risks to human health due to its neurotoxicity, endocrine disruption, and reproductive toxicity. Sensitive and rapid monitoring of carbendazim residues in food is essential for ensuring food safety. Among the numerous developed methods, electrochemical sensing stands out due to its simplicity of operation, low cost, and capability for on-site detection. Its key point is to construct a high-performance sensing interface. Nevertheless, these interfaces still face some limitations, such as inadequate conductivity, relatively low sensitivity, and narrow linear range.
Results
Herein, graphene supported iron nanoparticles encapsulated nitrogen-doped carbon nanotubes (Fe@N-CNTs) were constructed. The MOF-derived “tube-bridge” CNTs architecture, enhanced by N-doping, ensures high conductivity and optimized electron transfer pathways. Encapsulated iron nanoparticles act as shielded yet accessible redox-active centers, enhancing catalytic specificity. Graphene prevents CNTs stacking, maximizing active site exposure and interfacial charge transfer. This synergistic design significantly boosts the electrochemical active surface area, electron transfer kinetics, and electrocatalytic activity. As a result, a highly sensitive electrochemical sensing platform for CBZ has been successfully constructed. An irreversible two-electron and two-proton process takes place on the surface of Fe@N-CNTs/Graphene/GCE. The composite demonstrated exceptional performance for CBZ detection, achieving a broad linear range (0.001–7.0 μM), and an ultralow detection limit (0.56 nM). Furthermore, the sensor exhibited superior repeatability, stability, and selectivity. Its practical applicability was successfully validated through the accurate quantification of CBZ residues in medicinal herbs and vegetables (recoveries of 98.58 %–101.7 %).
Significance
This work offering a robust solution for monitoring ultra-low concentrations CBZ residues in complex food matrices. This innovative technology provides essential support for combating pesticide misuse while safeguarding public health and preserving ecological integrity related to CBZ contamination.
{"title":"Fe-doped ZnMOF-derived heterocarbon nanotubes for electrochemical sensing of carbendazim residues in food","authors":"Yicheng Yao , Liudi Ji , Peng Hu , Xiaoming Zhu , Junxing Hao , Kangbing Wu","doi":"10.1016/j.aca.2026.345103","DOIUrl":"10.1016/j.aca.2026.345103","url":null,"abstract":"<div><h3>Background</h3><div>Carbendazim (CBZ), is a benzimidazole fungicide and widely used for controlling fruit and vegetable diseases, which poses significant risks to human health due to its neurotoxicity, endocrine disruption, and reproductive toxicity. Sensitive and rapid monitoring of carbendazim residues in food is essential for ensuring food safety. Among the numerous developed methods, electrochemical sensing stands out due to its simplicity of operation, low cost, and capability for on-site detection. Its key point is to construct a high-performance sensing interface. Nevertheless, these interfaces still face some limitations, such as inadequate conductivity, relatively low sensitivity, and narrow linear range.</div></div><div><h3>Results</h3><div>Herein, graphene supported iron nanoparticles encapsulated nitrogen-doped carbon nanotubes (Fe@N-CNTs) were constructed. The MOF-derived “tube-bridge” CNTs architecture, enhanced by N-doping, ensures high conductivity and optimized electron transfer pathways. Encapsulated iron nanoparticles act as shielded yet accessible redox-active centers, enhancing catalytic specificity. Graphene prevents CNTs stacking, maximizing active site exposure and interfacial charge transfer. This synergistic design significantly boosts the electrochemical active surface area, electron transfer kinetics, and electrocatalytic activity. As a result, a highly sensitive electrochemical sensing platform for CBZ has been successfully constructed. An irreversible two-electron and two-proton process takes place on the surface of Fe@N-CNTs/Graphene/GCE. The composite demonstrated exceptional performance for CBZ detection, achieving a broad linear range (0.001–7.0 μM), and an ultralow detection limit (0.56 nM). Furthermore, the sensor exhibited superior repeatability, stability, and selectivity. Its practical applicability was successfully validated through the accurate quantification of CBZ residues in medicinal herbs and vegetables (recoveries of 98.58 %–101.7 %).</div></div><div><h3>Significance</h3><div>This work offering a robust solution for monitoring ultra-low concentrations CBZ residues in complex food matrices. This innovative technology provides essential support for combating pesticide misuse while safeguarding public health and preserving ecological integrity related to CBZ contamination.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345103"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145993127","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-22Epub Date: 2026-01-13DOI: 10.1016/j.aca.2026.345082
Qing'ai Chen , Yuwei Zhou , Guobin Huang , Chenchen Lin , Xucong Lin
Background
Shellfish toxin residues (e.g., okadaic acid, OA) in seafood are a global concern, and the development of precise, sensitive portable test kits is therefore necessary. Magnetic nanoparticle-aptasensors (MNAS) are attractive for specific recognition and sensitive assay, yet direct epitaxial aptamer immobilization on Fe3O4 causes thin, scattered coatings, leading to limited probe capacity and non-specific adsorption. Fabricating high-efficiency MNAS with dense, ordered aptamer arrays for precise detection thus remains a challenging yet promising goal.
Results
We propose a high-performance magnetic nanoparticle-aptasensor (MNAS) mediated by UiO-66-NH2 MOFs prepared with a secondary growth strategy (denoted as UiO/UiO-MOFs), in which AuNPs were uniformly anchored on MOF and facilitated the orientation-ordered nucleic acid aptamers enabling specific test of trace OA. The UiO/UiO-MOFs served as a scaffold with ordered amino group arrays, facilitating the construction of a homogeneous AuNPs interface and thus the tethering of high-density aptamers (up to 12.5 μmol/kg). This sensing platform synergistically integrated the advantages of ordered UiO/UiO-MOF self-assembly, robust AuNPs bridging, and the spherically ordered configuration of dense aptamers, thereby achieving efficient target OA recognition with minimal non-specific cross-reactivity. Coupled with laser-induced fluorescence (LIF) detection, the sensor exhibited excellent specificity, ultra-high sensitivity (limit of detection, LOD = 30 pg/mL), and outstanding stability. Notably, the sensor was successfully applied for the accurate on-site detection of OA residues in real mussel and clam samples, with satisfactory recovery rates ranging from 83.0 ± 3.3 % to 107.7 ± 1.3 %.
Significance
This study posed an affordable yet powerful strategy capable of simultaneously realizing efficient UiO/UiO-MOFs surface mediation surface mediation and high capacity of spherical orientation-ordered aptamers. It expands the toolkit for epitaxial aptamer immobilization on MNAS, facilitates the ordered anchoring of aptamers, and thus enables the construction of advanced MNAS tailored for shellfish toxin detection.
{"title":"Magnetic aptasensor with UiO-MOFs-mediated orientation-ordered aptamers for sensitive portable test of okadaic acid toxin","authors":"Qing'ai Chen , Yuwei Zhou , Guobin Huang , Chenchen Lin , Xucong Lin","doi":"10.1016/j.aca.2026.345082","DOIUrl":"10.1016/j.aca.2026.345082","url":null,"abstract":"<div><h3>Background</h3><div>Shellfish toxin residues (e.g., okadaic acid, OA) in seafood are a global concern, and the development of precise, sensitive portable test kits is therefore necessary. Magnetic nanoparticle-aptasensors (MNAS) are attractive for specific recognition and sensitive assay, yet direct epitaxial aptamer immobilization on Fe<sub>3</sub>O<sub>4</sub> causes thin, scattered coatings, leading to limited probe capacity and non-specific adsorption. Fabricating high-efficiency MNAS with dense, ordered aptamer arrays for precise detection thus remains a challenging yet promising goal.</div></div><div><h3>Results</h3><div>We propose a high-performance magnetic nanoparticle-aptasensor (MNAS) mediated by UiO-66-NH<sub>2</sub> MOFs prepared with a secondary growth strategy (denoted as UiO/UiO-MOFs), in which AuNPs were uniformly anchored on MOF and facilitated the orientation-ordered nucleic acid aptamers enabling specific test of trace OA. The UiO/UiO-MOFs served as a scaffold with ordered amino group arrays, facilitating the construction of a homogeneous AuNPs interface and thus the tethering of high-density aptamers (up to 12.5 μmol/kg). This sensing platform synergistically integrated the advantages of ordered UiO/UiO-MOF self-assembly, robust AuNPs bridging, and the spherically ordered configuration of dense aptamers, thereby achieving efficient target OA recognition with minimal non-specific cross-reactivity. Coupled with laser-induced fluorescence (LIF) detection, the sensor exhibited excellent specificity, ultra-high sensitivity (limit of detection, LOD = 30 pg/mL), and outstanding stability. Notably, the sensor was successfully applied for the accurate on-site detection of OA residues in real mussel and clam samples, with satisfactory recovery rates ranging from 83.0 ± 3.3 % to 107.7 ± 1.3 %.</div></div><div><h3>Significance</h3><div>This study posed an affordable yet powerful strategy capable of simultaneously realizing efficient UiO/UiO-MOFs surface mediation surface mediation and high capacity of spherical orientation-ordered aptamers. It expands the toolkit for epitaxial aptamer immobilization on MNAS, facilitates the ordered anchoring of aptamers, and thus enables the construction of advanced MNAS tailored for shellfish toxin detection.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345082"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145957110","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-22Epub Date: 2026-01-13DOI: 10.1016/j.aca.2026.345098
Mikhail I. Bogachev , Pavel S. Baranov , Aleksandr M. Sinitca , Anna V. Mironova , Dmitry R. Sharivzyanov , Alexander A. Basmanov , Elena Y. Trizna , Anna S. Gorshkova , Nikita S. Pyko , Airat R. Kayumov
Background
Biofilms are present on almost all surfaces in households, healthcare and medical equipment, foods, industrial and water supply systems, and often contain opportunistic pathogens that represent one of the key public health hazards. The highest risks are imposed by ESKAPEE pathogens, as well as mixed bacterial-fungal communities often exhibiting multiple drug resistance, this way challenging public healthcare.
Results
Here we show how recent developments in hyperspectral imaging technology, complemented by advanced image analysis and machine learning methods, create a unique framework for the potential design of non-contact monitoring systems operating at the macroscale. We could successfully identify five key pathogenic bacteria and a common pathogenic yeast, C. albicans, that frequently occur on surfaces in monospecies and mixed biofilms consisting of combinations of various strains using hyperspectral imaging in the visible, near-infrared, and short-wave infrared spectral bands. Our results indicate that the above pathogenic species could be identified in monocultural biofilms with 95–99.5 % accuracy, while in more frequently occurring mixed biofilms obtained by combining different microorganisms, the accuracy ranges from 90 to 92 % for gram-negative E. coli, K. pneumoniae, and P. aeruginosa to 96–99 % for fungi and gram-positive E. faecalis and S. aureus, respectively, under in vitro conditions.
Significance
Based on our results, we believe that hyperspectral imaging represents a promising and highly efficient technology for the rapid, regular, non-contact monitoring of various equipment and surfaces to detect bacterial and fungal pathogens in situ.
{"title":"Non-contact identification of opportunistic pathogens in mixed biofilm contaminations by hyperspectral imaging","authors":"Mikhail I. Bogachev , Pavel S. Baranov , Aleksandr M. Sinitca , Anna V. Mironova , Dmitry R. Sharivzyanov , Alexander A. Basmanov , Elena Y. Trizna , Anna S. Gorshkova , Nikita S. Pyko , Airat R. Kayumov","doi":"10.1016/j.aca.2026.345098","DOIUrl":"10.1016/j.aca.2026.345098","url":null,"abstract":"<div><h3>Background</h3><div>Biofilms are present on almost all surfaces in households, healthcare and medical equipment, foods, industrial and water supply systems, and often contain opportunistic pathogens that represent one of the key public health hazards. The highest risks are imposed by ESKAPEE pathogens, as well as mixed bacterial-fungal communities often exhibiting multiple drug resistance, this way challenging public healthcare.</div></div><div><h3>Results</h3><div>Here we show how recent developments in hyperspectral imaging technology, complemented by advanced image analysis and machine learning methods, create a unique framework for the potential design of non-contact monitoring systems operating at the macroscale. We could successfully identify five key pathogenic bacteria and a common pathogenic yeast, <em>C. albicans,</em> that frequently occur on surfaces in monospecies and mixed biofilms consisting of combinations of various strains using hyperspectral imaging in the visible, near-infrared, and short-wave infrared spectral bands. Our results indicate that the above pathogenic species could be identified in monocultural biofilms with 95–99.5 % accuracy, while in more frequently occurring mixed biofilms obtained by combining different microorganisms, the accuracy ranges from 90 to 92 % for gram-negative <em>E. coli</em>, <em>K. pneumoniae,</em> and <em>P. aeruginosa</em> to 96–99 % for fungi and gram-positive <em>E. faecalis</em> and <em>S. aureus</em>, respectively, under <em>in vitro</em> conditions.</div></div><div><h3>Significance</h3><div>Based on our results, we believe that hyperspectral imaging represents a promising and highly efficient technology for the rapid, regular, non-contact monitoring of various equipment and surfaces to detect bacterial and fungal pathogens <em>in situ</em>.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1388 ","pages":"Article 345098"},"PeriodicalIF":6.0,"publicationDate":"2026-02-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145955918","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-15Epub Date: 2025-11-24DOI: 10.1016/j.aca.2025.344932
Mengru Xie , Jinghui Ma , Xiaoxing Zhou , Yan Wei , Yilian Zhao , Chao Ye , Xinchu Liu , Jilin Qing , Zhizhong Chen
Recombinase-Aided Amplification (RAA), as an emerging isothermal nucleic acid amplification technique, demonstrates significant potential in infectious disease diagnostics due to its advantages of rapidity, high efficiency, and reduced reliance on complex instrumentation. Amid frequent global outbreaks of infectious diseases, the demand for rapid and accurate diagnostic technologies has intensified. RAA, leveraging its unique amplification mechanism and application flexibility, has garnered increasing research interest. Compared to other amplification techniques, RAA exhibits shorter reaction times and higher sensitivity, enabling the detection of diverse pathogens including viruses, bacteria, fungi, and parasites. Furthermore, the integration of RAA with techniques such as CRISPR and microfluidics provides innovative strategies for point-of-care testing. However, current RAA implementation faces industrial bottlenecks such as low primer design tolerance, nonspecific amplification due to sample matrix interference, and lack of standardized frameworks. This review aims to summarize the fundamental principles of RAA technology, its current applications in infectious disease diagnosis, synergistic innovations with complementary methods, and future development trajectories, thereby offering theoretical insights for the optimization and advancement of rapid diagnostic technologies.
{"title":"Recombinase-aided amplification technology: A comprehensive review of innovations and multi-technology integration for rapid infectious disease diagnosis","authors":"Mengru Xie , Jinghui Ma , Xiaoxing Zhou , Yan Wei , Yilian Zhao , Chao Ye , Xinchu Liu , Jilin Qing , Zhizhong Chen","doi":"10.1016/j.aca.2025.344932","DOIUrl":"10.1016/j.aca.2025.344932","url":null,"abstract":"<div><div>Recombinase-Aided Amplification (RAA), as an emerging isothermal nucleic acid amplification technique, demonstrates significant potential in infectious disease diagnostics due to its advantages of rapidity, high efficiency, and reduced reliance on complex instrumentation. Amid frequent global outbreaks of infectious diseases, the demand for rapid and accurate diagnostic technologies has intensified. RAA, leveraging its unique amplification mechanism and application flexibility, has garnered increasing research interest. Compared to other amplification techniques, RAA exhibits shorter reaction times and higher sensitivity, enabling the detection of diverse pathogens including viruses, bacteria, fungi, and parasites. Furthermore, the integration of RAA with techniques such as CRISPR and microfluidics provides innovative strategies for point-of-care testing. However, current RAA implementation faces industrial bottlenecks such as low primer design tolerance, nonspecific amplification due to sample matrix interference, and lack of standardized frameworks. This review aims to summarize the fundamental principles of RAA technology, its current applications in infectious disease diagnosis, synergistic innovations with complementary methods, and future development trajectories, thereby offering theoretical insights for the optimization and advancement of rapid diagnostic technologies.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1387 ","pages":"Article 344932"},"PeriodicalIF":6.0,"publicationDate":"2026-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145583709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-15Epub Date: 2026-01-07DOI: 10.1016/j.aca.2026.345071
Jie Zhang , Jinna Xiao , Zhengzheng Liao , Haijiao Xie , Zhentao Li
Background
Quetiapine (QTP) and clozapine (CLZ) are widely used antipsychotic drugs to treat psychotic symptoms. However, their narrow therapeutic window and the potential risk of severe side effects necessitate close monitoring of blood concentrations. Current therapeutic drug monitoring (TDM) methods are still limited by operational complexity, loss of trace components, and potential instrument contamination. This study proposed a solid-phase microextraction (SPME) fiber coated with a multi-component covalent organic framework (COF TAB-DHTA-DMTP), aiming to achieve highly efficient extraction of psychiatric drugs in human plasma.
Results
The COF TAB-DHTA-DMTP, characterized by a large specific surface area and well-defined porosity, was embedded in polyacrylonitrile (PAN) to produce a matrix-compatible coating material for SPME, which was subsequently coupled with high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) for the determination of QTP and CLZ. Under the optimal SPME parameters, the developed method exhibited excellent linearity for QTP and CLZ within the concentration range of 1–1000 ng/mL, ranging from 0.9994 to 0.9996, low limits of detection (LODs) between 0.021 and 0.035 ng/mL, and satisfactory precision, with relative standard deviations (RSDs) below 5.10 %. The extraction mechanism was elucidated through theoretical simulations and experimental characterizations. Additionally, clinical samples obtained from hospitals were analyzed using the proposed SPME method, and the results were validated against those obtained by conventional laboratory-based method.
Significance and novelty
This novel self-fabricated COF-based SPME fiber can effectively extract QTP and CLZ from complex human plasma samples, eliminating interfering substances, mitigating matrix effects, enhancing detection sensitivity, and reducing the risk of instrument contamination. Our research has facilitated the application of SPME technology from laboratory settings to clinical practice, further confirming its applicability in the clinical monitoring of psychiatric drugs.
{"title":"Construction of multi-component covalent organic framework coated stainless steel fiber for efficient microextraction of psychiatric drugs: experiments and clinical applications","authors":"Jie Zhang , Jinna Xiao , Zhengzheng Liao , Haijiao Xie , Zhentao Li","doi":"10.1016/j.aca.2026.345071","DOIUrl":"10.1016/j.aca.2026.345071","url":null,"abstract":"<div><h3>Background</h3><div>Quetiapine (QTP) and clozapine (CLZ) are widely used antipsychotic drugs to treat psychotic symptoms. However, their narrow therapeutic window and the potential risk of severe side effects necessitate close monitoring of blood concentrations. Current therapeutic drug monitoring (TDM) methods are still limited by operational complexity, loss of trace components, and potential instrument contamination. This study proposed a solid-phase microextraction (SPME) fiber coated with a multi-component covalent organic framework (COF TAB-DHTA-DMTP), aiming to achieve highly efficient extraction of psychiatric drugs in human plasma.</div></div><div><h3>Results</h3><div>The COF TAB-DHTA-DMTP, characterized by a large specific surface area and well-defined porosity, was embedded in polyacrylonitrile (PAN) to produce a matrix-compatible coating material for SPME, which was subsequently coupled with high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) for the determination of QTP and CLZ. Under the optimal SPME parameters, the developed method exhibited excellent linearity for QTP and CLZ within the concentration range of 1–1000 ng/mL, ranging from 0.9994 to 0.9996, low limits of detection (LODs) between 0.021 and 0.035 ng/mL, and satisfactory precision, with relative standard deviations (RSDs) below 5.10 %. The extraction mechanism was elucidated through theoretical simulations and experimental characterizations. Additionally, clinical samples obtained from hospitals were analyzed using the proposed SPME method, and the results were validated against those obtained by conventional laboratory-based method.</div></div><div><h3>Significance and novelty</h3><div>This novel self-fabricated COF-based SPME fiber can effectively extract QTP and CLZ from complex human plasma samples, eliminating interfering substances, mitigating matrix effects, enhancing detection sensitivity, and reducing the risk of instrument contamination. Our research has facilitated the application of SPME technology from laboratory settings to clinical practice, further confirming its applicability in the clinical monitoring of psychiatric drugs.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1387 ","pages":"Article 345071"},"PeriodicalIF":6.0,"publicationDate":"2026-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145914904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-15Epub Date: 2026-01-07DOI: 10.1016/j.aca.2026.345075
Yang Liu , Xiaorun Qi , Qian Sun , Manman Wang , Yu He , Gongwu Song
Background
N-phenyl-N′-(1,3-dimethylbutyl)-p-phenylenediamine-quinone (6PPD-Q), an emerging pollutant, is a highly toxic chemical derived from tires, which have possible adverse effects on human health via the food chain. Despite the widespread occurrence of 6PPD-Q in the environment, methods for its detection remain relatively scarce. Consequently, the development of accurate methods for the quick, highly responsive, and specific analysis of 6PPD-Q is critical.
Results
A deep learning and smartphone integrated ratiometric fluorescence sensor was proposed for the point-of-care testing (POCT) of 6PPD-Q. Silicon quantum dots (SiQDs) and [Ru(bpy)2(phen-NH2)]2+ were employed for preparing the SiQDs@[Ru(bpy)2(phen-NH2)]2+ complex with two emissions at 430 nm and 610 nm respectively, which corresponds to blue and red fluorescence colors. As increased concentration of 6PPD-Q, the fluorescence of SiQDs@[Ru(bpy)2(phen-NH2)]2+ complex at 430 nm was gradually enhanced, while the fluorescence at 610 nm kept unchanged. When the concentration of 6PPD-Q was ranging from 0.0066 to 2 μg mL−1 and 2–7 μg mL−1, the fluorescence intensity ratio I430/I610 and the concentration of 6PPD-Q gave an outstanding linear relationship with the limit of detection (LOD) of 2 ng mL−1. Meanwhile, the fluorescence color change of SiQDs@[Ru(bpy)2(phen-NH2)]2+ underwent a gradual transition from red to magenta and finally to purple, which could be recognized by the naked eye. The YOLOv5 algorithm was utilized to locate the fluorescence images and acquired the corresponding RGB values for deep learning to enhance the accuracy.
Significance
This study established an analytical platform for the analysis of 6PPD-Q with the merits of low cost, convenience, visualization and real-time detection, which is expected to open a new avenue for the detection of emerging pollutants.
n -苯基- n ' -（1,3-二甲基丁基）-对苯二胺醌（6PPD-Q）是一种来自轮胎的剧毒化学物质，可能通过食物链对人体健康产生不利影响，是一种新兴污染物。尽管6PPD-Q在环境中广泛存在，但其检测方法仍然相对缺乏。因此，开发准确的方法来快速、高响应和特异性分析6PPD-Q是至关重要的。结果提出了一种深度学习智能手机集成比例荧光传感器用于6PPD-Q的即时检测（POCT）。采用硅量子点（SiQDs）和[Ru(bpy)2(phenn - nh2)]2+制备了SiQDs@[Ru(bpy)2(phenn - nh2)]2+配合物，在430 nm和610 nm处分别发出蓝色和红色荧光。随着6PPD-Q浓度的增加，SiQDs@[Ru(bpy)2(phenn - nh2)]2+配合物在430 nm处的荧光逐渐增强，而在610 nm处的荧光保持不变。6PPD-Q浓度在0.0066 ~ 2 μg mL−1和2 ~ 7 μg mL−1范围内，荧光强度比I430/I610与6PPD-Q浓度呈良好的线性关系，检测限（LOD）为2 ng mL−1。同时SiQDs@[Ru(bpy)2(phenn - nh2)]2+的荧光颜色变化经历了从红色到品红，最后到紫色的渐变过程，可以用肉眼识别。利用YOLOv5算法对荧光图像进行定位，获取相应的RGB值进行深度学习，提高准确率。本研究建立了一个成本低、方便、可视化、实时检测的6PPD-Q分析平台，有望为新兴污染物的检测开辟新的途径。
{"title":"SiQDs and [Ru(bpy)2(phen-NH2)]2+ based ratiometric fluorescence probe for point-of-care testing of 6PPD-quinone with 3D-printing portable devices","authors":"Yang Liu , Xiaorun Qi , Qian Sun , Manman Wang , Yu He , Gongwu Song","doi":"10.1016/j.aca.2026.345075","DOIUrl":"10.1016/j.aca.2026.345075","url":null,"abstract":"<div><h3>Background</h3><div>N-phenyl-N′-(1,3-dimethylbutyl)-p-phenylenediamine-quinone (6PPD-Q), an emerging pollutant, is a highly toxic chemical derived from tires, which have possible adverse effects on human health via the food chain. Despite the widespread occurrence of 6PPD-Q in the environment, methods for its detection remain relatively scarce. Consequently, the development of accurate methods for the quick, highly responsive, and specific analysis of 6PPD-Q is critical.</div></div><div><h3>Results</h3><div>A deep learning and smartphone integrated ratiometric fluorescence sensor was proposed for the point-of-care testing (POCT) of 6PPD-Q. Silicon quantum dots (SiQDs) and [Ru(bpy)<sub>2</sub>(phen-NH<sub>2</sub>)]<sup>2+</sup> were employed for preparing the SiQDs@[Ru(bpy)<sub>2</sub>(phen-NH<sub>2</sub>)]<sup>2+</sup> complex with two emissions at 430 nm and 610 nm respectively, which corresponds to blue and red fluorescence colors. As increased concentration of 6PPD-Q, the fluorescence of SiQDs@[Ru(bpy)<sub>2</sub>(phen-NH<sub>2</sub>)]<sup>2+</sup> complex at 430 nm was gradually enhanced, while the fluorescence at 610 nm kept unchanged. When the concentration of 6PPD-Q was ranging from 0.0066 to 2 μg mL<sup>−1</sup> and 2–7 μg mL<sup>−1</sup>, the fluorescence intensity ratio I<sub>430</sub>/I<sub>610</sub> and the concentration of 6PPD-Q gave an outstanding linear relationship with the limit of detection (LOD) of 2 ng mL<sup>−1</sup>. Meanwhile, the fluorescence color change of SiQDs@[Ru(bpy)<sub>2</sub>(phen-NH<sub>2</sub>)]<sup>2+</sup> underwent a gradual transition from red to magenta and finally to purple, which could be recognized by the naked eye. The YOLOv5 algorithm was utilized to locate the fluorescence images and acquired the corresponding RGB values for deep learning to enhance the accuracy.</div></div><div><h3>Significance</h3><div>This study established an analytical platform for the analysis of 6PPD-Q with the merits of low cost, convenience, visualization and real-time detection, which is expected to open a new avenue for the detection of emerging pollutants.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1387 ","pages":"Article 345075"},"PeriodicalIF":6.0,"publicationDate":"2026-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145915396","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2026-02-15Epub Date: 2026-01-08DOI: 10.1016/j.aca.2026.345072
Bingyu Peng , Qiaohui Zhong , Feiyu Dong , Gaobin Chu , Lu Yin , Jing Zhang , Jie Li
Background
Molybdenum (Mo) isotope has been widely used to trace various redox processes in earth science, however, conventional separation methods, typically depend on toxic hydrofluoric acid (HF), redox reagents (e.g., hydrogen peroxide (H2O2) and ascorbic acid (HAsc)), or multi-column purification steps, which introduce either safety hazards or operational complexity. In this study, a safe and efficient single-column procedure without use of either HF or redox reagents was developed for Mo separation from various samples for high precision Mo isotope measurements.
Results
This method takes advantage of high selective adsorption of Mo on an in-house TOPO resin and only utilize the diluted HCl - HNO3 media, achieving an effective matrix removal, a high Mo recovery (>93 %) and low procedural blanks (<0.50 ng). Repeated analyses of NIST SRM 3134, Mo metal, and seawater CASS-6 yielded an external reproducibility better than ±0.06 ‰ (2SD). Mn-doping experiments revealed that Mn/Mo ratios exceeding 0.50 induced significant polyatomic interference, causing measurable negative shifts (>0.10 ‰) in δ98/95MoNIST SRM 3134 values. The method was validated using 11 geological reference materials, showing good agreement with published values. Additionally, the δ98/95MoNIST SRM 3134 values of five biological reference materials (e.g., green tea, human hair, pig liver, and fetal bovine serum) were reported for the first time, exhibiting a significant isotopic variation of 1.44 ‰.
Significance and novelty
Compared with anion/cation-exchange resins or extraction resins, this novel procedure based on in-house TOPO resin depends on dilute HCl - HNO3 media and effectively purifies Mo from the matrix, with good performance and operational safety. Additionally, the δ98/95MoNIST SRM 3134 values for biological reference materials provide valuable references for quality assurance and inter-laboratory comparisons in emerging applications of Mo isotopes in biological systems. Overall, this study provided a simpler and safer alternative method for high-precise determination of Mo isotope in various samples.
{"title":"A novel and efficient single column separation procedure for high-precision determination of Molybdenum isotope ratio analysis by MC-ICP-MS","authors":"Bingyu Peng , Qiaohui Zhong , Feiyu Dong , Gaobin Chu , Lu Yin , Jing Zhang , Jie Li","doi":"10.1016/j.aca.2026.345072","DOIUrl":"10.1016/j.aca.2026.345072","url":null,"abstract":"<div><h3>Background</h3><div>Molybdenum (Mo) isotope has been widely used to trace various redox processes in earth science, however, conventional separation methods, typically depend on toxic hydrofluoric acid (HF), redox reagents (e.g., hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) and ascorbic acid (HAsc)), or multi-column purification steps, which introduce either safety hazards or operational complexity. In this study, a safe and efficient single-column procedure without use of either HF or redox reagents was developed for Mo separation from various samples for high precision Mo isotope measurements.</div></div><div><h3>Results</h3><div>This method takes advantage of high selective adsorption of Mo on an in-house TOPO resin and only utilize the diluted HCl - HNO<sub>3</sub> media, achieving an effective matrix removal, a high Mo recovery (>93 %) and low procedural blanks (<0.50 ng). Repeated analyses of NIST SRM 3134, Mo metal, and seawater CASS-6 yielded an external reproducibility better than ±0.06 ‰ (2SD). Mn-doping experiments revealed that Mn/Mo ratios exceeding 0.50 induced significant polyatomic interference, causing measurable negative shifts (>0.10 ‰) in δ<sup>98/95</sup>Mo<sub>NIST SRM 3134</sub> values. The method was validated using 11 geological reference materials, showing good agreement with published values. Additionally, the δ<sup>98/95</sup>Mo<sub>NIST SRM 3134</sub> values of five biological reference materials (e.g., green tea, human hair, pig liver, and fetal bovine serum) were reported for the first time, exhibiting a significant isotopic variation of 1.44 ‰.</div></div><div><h3>Significance and novelty</h3><div>Compared with anion/cation-exchange resins or extraction resins, this novel procedure based on in-house TOPO resin depends on dilute HCl - HNO<sub>3</sub> media and effectively purifies Mo from the matrix, with good performance and operational safety. Additionally, the δ<sup>98/95</sup>Mo<sub>NIST SRM 3134</sub> values for biological reference materials provide valuable references for quality assurance and inter-laboratory comparisons in emerging applications of Mo isotopes in biological systems. Overall, this study provided a simpler and safer alternative method for high-precise determination of Mo isotope in various samples.</div></div>","PeriodicalId":240,"journal":{"name":"Analytica Chimica Acta","volume":"1387 ","pages":"Article 345072"},"PeriodicalIF":6.0,"publicationDate":"2026-02-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145920096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
扫码关注我们
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号