Researchers have shown significant interest in modulating the peroxidase-like activity of nanozymes.Among these,bimetallic nanozymes have shown superior peroxidase-like activity over monometallic counterparts,offering...Researchers have shown significant interest in modulating the peroxidase-like activity of nanozymes.Among these,bimetallic nanozymes have shown superior peroxidase-like activity over monometallic counterparts,offering enhanced performance and cost-efficiency in nanozyme designs.Herein,bimetallic nanozymes comprising nickel(Ni)and osmium(Os)incorporated into hyaluronate(HA)have been developed,resulting in HA-Nin/Os nanoclusters.Subsequently,comprehensive characterizations have been conducted.Further investigation has revealed that HA-Nin/Os efficiently catalyzed 3,3,5,5-tetramethylbenzidine(TMB)oxidation with hydrogen peroxide(H_(2)O_(2)),confirming its peroxidase-like behavior and role as a nanozyme.Impressively,HA-Ni_(2)/Os(Ni/Os=2:1)displays heightened substrate affinity,accelerated reaction rates,enhanced hydroxyl radical production in acidic conditions,and exhibits activity unit of 1224 U/mg,representing more than two-fold increase compared to non-Ni-supported Os nanozyme.Theoretical calculations indicate that Ni support enhances the peroxidase-like process of Os nanozyme by improving H_(2)O_(2) adsorption and TMB oxidation.Crucially,the support of Ni does not significantly alter the other enzyme-like activities of Os nanozymes,thereby enabling Ni to selectively enhance their peroxidase-like activity.In terms of application,the peroxidase-like ability of HA-Ni_(2)/Os,facilitated by HA's carboxyl groups enabling crosslinking,proves effective in a squamous carcinoma antigen immunoassay.Moreover,HA-Ni_(2)/Os exhibit reliable stability,promising as a peroxidase substitute.This work underscores the advantages of incorporating Ni into Os,specifically enhancing peroxidase-like activity,highlighting the potential of Os bimetallic nanozymes for peroxidase-based applications.展开更多
Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitori...Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitoring.Nevertheless,challenges still exist in detecting ultra-low concentration analytes due to the inherent sensitivity limitations of LFIA.Recently,significant advances have been achieved by integrating enzyme activity probes and transforming LFIA into a highly sensitive tool for rapidly detecting trace analyte concentrations.Specifically,modifying natural enzymes or engineered nanozymes allows them to function as immune probes,directly catalyzing the production of signal molecules or indirectly initiating enzyme activity.Therefore,the signal intensity and detection sensitivity of LFIA are markedly elevated.The present review undertakes a comprehensive examination of pertinent research literature,offering a systematic analysis of recently proposed enzyme-based signal amplification strategies.By way of comparative assessment,the merits and demerits of current approaches are delineated,along with the identification of research avenues that still need to be explored.It is anticipated that this critical overview will garner considerable attention within the biomedical and materials science communities,providing valuable direction and insight toward the advancement of high-performance LFIA technologies.展开更多
The advancement of various types of fluorescent nanoparticles is crucial for enhancing the application of lateral flow immunoassays(LFIA)across multiple fields.Currently,the fluorescent nanoparticles utilized in LFIA ...The advancement of various types of fluorescent nanoparticles is crucial for enhancing the application of lateral flow immunoassays(LFIA)across multiple fields.Currently,the fluorescent nanoparticles utilized in LFIA predominantly consist of traditional dye-doped nanoparticles or aggregation-induced luminescence dye-doped nanoparticles.The reliance on specific types of nanoparticles limits the diversity of signal reporting groups available for LFIA.Herein,we developed a solid-state luminescent dye-doped nanoparticles(SLDNPs)-based LFIA system with exceptional stability for the detection of C-reactive protein(CRP)in serum.The synthesis of SLD_(520)NP_(S)was simplicity,efficient and eco-friendly,which was ideal for large-scale production of the LFIA test strip.And the SLD_(520)NP_(S)exhibits superior fluorescence quantum yield(49%),fully guarantees the performance of the LFIA test strip.The constructed SLD_(520)NPsm Ab1-based LFIA demonstrated a satisfactory linear relationship with CRP concentrations ranging from 0.5 ng/mL to 100 ng/mL,with limits of detection(LOD)of 0.78 ng/mL and a visible LOD of 1 ng/mL using a handheld 405 nm lamp.Furthermore,the developed LFIA exhibited excellent recoveries in serum,ranging from 94.45%to 102.5%.Overall,the outstanding performance of the SLD_(520)NPs-mAb1-based LFIA indicates that solid-state luminescent dyes have significant potential applications in the field of LFIA.展开更多
Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,...Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,we reported a liposome-mediated signal-off photoelectrochemical(PEC)immunoassay for the sensitive detection of carcinoembryonic antigen(CEA)using ternary transition metal sulfide CuS/ZnCdS as the photoactive material.Good photocurrents were acquired on the basis of specific oxidation reaction of dopamine on the CuS/ZnCdS.The energy band relationship of CuS/ZnCdS was determined,and the wellmatched oxidation potential of dopamine was verified.To achieve accurate recovery of low-abundance CEA,systematic PEC evaluation from human serum samples was performed by combining with classical immunoreaction and liposome-induced dopamine amplification strategy with high stability and selectivity.Under optimum conditions,PEC immunoassay displayed good photocurrent responses toward target CEA with a dynamic linear range of 0.1-50 ng/mL with a detection limit of 31.6 pg/mL.Importantly,this system by combining with a discussion of energy level matching between semiconductor energy bands and small-molecules opens a new horizon for development of high-efficient PEC immunoassays.展开更多
This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to c...This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to compare its performance with the chemiluminescence microparticle immunoassay(CMIA).A total of 211 serum samples from pediatric patients with intracranial tumors undergoing high-dose MTX treatment were analyzed using both techniques.Correlation and agreement analyses were performed to assess the level of concordance between these methods.The results demonstrated a significant correlation(P<0.05)between the two methods,with correlation coefficients of 0.9913 and 0.9893,respectively.However,a statistical difference was noted in MTX serum concentrations at lower levels(0.04-1.5μmol/L),while no significant difference was observed at higher concentrations(1.5-400μmol/L).Specifically,in the 0.04-1.5μmol/L range(107 cases),Bland-Altman analysis indicated a bias of 0.018 between the two methods.Given the observed discrepancies,particularly at lower concentrations,it is advised that the UPLC-MS/MS method should not be used interchangeably with the CMIA assay for therapeutic drug monitoring in patients receiving high-dose MTX treatment.展开更多
We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBO...We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.展开更多
The application of pesticides (mostly insecticides and fungicides) during the tea-planting process will undoubtedly increase the dietary risk associated with drinking tea. Thus, it is necessary to ascertain whether pe...The application of pesticides (mostly insecticides and fungicides) during the tea-planting process will undoubtedly increase the dietary risk associated with drinking tea. Thus, it is necessary to ascertain whether pesticide residues in tea products exceed the maximum residue limits. However, the complex matrices present in tea samples comprise a major challenge in the analytical detection of pesticide residues. In this study, nine types of lateral flow immunochromatographic strips (LFICSs) were developed to detect the pesticides of interest (fenpropathrin, chlorpyrifos, imidacloprid, thiamethoxam, acetamiprid, carbendazim, chlorothalonil, pyraclostrobin, and iprodione). To reduce the interference of tea substrates on the assay sensitivity, the pretreatment conditions for tea samples, including the extraction solvent, extraction time, and purification agent, were optimized for the simultaneous detection of these pesticides. The entire testing procedure (including pretreatment and detection) could be completed within 30 min. The detected results of authentic tea samples were confirmed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), which suggest that the LFICS coupled with sample rapid pretreatment can be used for on-site rapid screening of the target pesticide in tea products prior to their market release.展开更多
Foods are often contaminated by multiple foodborne pathogens,which threatens human health.In this work,we developed a microfluidic biosensor for multiplex immunoassay of foodborne bacteria with agitation driven by pro...Foods are often contaminated by multiple foodborne pathogens,which threatens human health.In this work,we developed a microfluidic biosensor for multiplex immunoassay of foodborne bacteria with agitation driven by programmed audio signals.This agitation,powered by the vibration of a speaker cone during music playing,accelerated the mass transport in the incubation process to form bacterial complexes within 10 min.Immunoassay reagents of the two target bacteria(Escherichia coli O157:H7 and Salmonella typhimurium)were preloaded into the corresponding fore-vacuum storage chamber on the chip,and released to participate in the subsequent immune analysis process by piercing the chambers.All the detection processes were integrated into a single microfluidic chip and controlled by a smartphone through Bluetooth.Under selected conditions,wide linear ranges and low limits of detection(LODs<2CFU/m L)were obtained,and real food samples were successfully determined within 30 min.This biosensing method can be extended to wide-ranging applications by loading different recognizing reagents.展开更多
[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PP...[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.展开更多
目的对人甲胎蛋白(alpha-fetoprotein,AFP)第2代国际标准品(编号:22/216)的含量进行协作标定研究。方法中国食品药品检定研究院代表中国实验室,组织了13家AFP试剂生产企业,采用13种化学发光免疫分析法、2种时间分辨荧光免疫法和1种酶联...目的对人甲胎蛋白(alpha-fetoprotein,AFP)第2代国际标准品(编号:22/216)的含量进行协作标定研究。方法中国食品药品检定研究院代表中国实验室,组织了13家AFP试剂生产企业,采用13种化学发光免疫分析法、2种时间分辨荧光免疫法和1种酶联免疫法的试剂盒,按照英国药品和健康产品管理局提供的统一研究方案,以AFP第1代国际标准品为标准标定22/216。结果中国实验室提交的22/216含量的几何平均值(geometric mean,GM)为7828.78IU/安瓿(95%CI:7462.49~8213.05IU/安瓿),其几何变异系数(geometric coefficient of variation,GCV)为9.77%(n=16)。全球4个国家9个实验室参加了本研究,全部数据得到的22/216含量GM为7756.21IU/安瓿(95%CI:7486.81~8035.30IU/安瓿),GCV为8.73%(n=24)。结论经世界卫生组织生物标准专家委员会审核,确定22/216可作为AFP第2代国际标准品使用,效价为7800IU/安瓿。展开更多
基金financial support from the Natural Science Foundation of Fujian Province(No.2022J01271)the Joint Funds for the Innovation of Science and Technology,Fujian Province(No.2023Y9226)+1 种基金the Introduced High-Level Talent Team Project of Quanzhou City(No.2023CT008)the Doctoral Research Foundation Project of the Second Affiliated Hospital of Fujian Medical University(No.BS202201)。
文摘Researchers have shown significant interest in modulating the peroxidase-like activity of nanozymes.Among these,bimetallic nanozymes have shown superior peroxidase-like activity over monometallic counterparts,offering enhanced performance and cost-efficiency in nanozyme designs.Herein,bimetallic nanozymes comprising nickel(Ni)and osmium(Os)incorporated into hyaluronate(HA)have been developed,resulting in HA-Nin/Os nanoclusters.Subsequently,comprehensive characterizations have been conducted.Further investigation has revealed that HA-Nin/Os efficiently catalyzed 3,3,5,5-tetramethylbenzidine(TMB)oxidation with hydrogen peroxide(H_(2)O_(2)),confirming its peroxidase-like behavior and role as a nanozyme.Impressively,HA-Ni_(2)/Os(Ni/Os=2:1)displays heightened substrate affinity,accelerated reaction rates,enhanced hydroxyl radical production in acidic conditions,and exhibits activity unit of 1224 U/mg,representing more than two-fold increase compared to non-Ni-supported Os nanozyme.Theoretical calculations indicate that Ni support enhances the peroxidase-like process of Os nanozyme by improving H_(2)O_(2) adsorption and TMB oxidation.Crucially,the support of Ni does not significantly alter the other enzyme-like activities of Os nanozymes,thereby enabling Ni to selectively enhance their peroxidase-like activity.In terms of application,the peroxidase-like ability of HA-Ni_(2)/Os,facilitated by HA's carboxyl groups enabling crosslinking,proves effective in a squamous carcinoma antigen immunoassay.Moreover,HA-Ni_(2)/Os exhibit reliable stability,promising as a peroxidase substitute.This work underscores the advantages of incorporating Ni into Os,specifically enhancing peroxidase-like activity,highlighting the potential of Os bimetallic nanozymes for peroxidase-based applications.
基金Financial supports from the National Natural Science Foundation of China(NSFC,Nos.52272144 and 22205048)Heilongjiang Provincial Natural Science Foundation of China(No.JQ2022E001)+3 种基金China Postdoctoral Science Foundation(Nos.2022M710931 and 2023T160154)Heilongjiang Postdoctoral Science Foundation(No.LBH-Z22010)Natural Science Foundation of Shandong Province(No.ZR2020ZD42)the Fundamental Research funds for the Central Universities are greatly acknowledged.
文摘Lateral flow immunoassay(LFIA),a rapid detection technique noted for simplicity and economy,has showcased indispensable applicability in diverse domains such as disease screening,food safety,and environmental monitoring.Nevertheless,challenges still exist in detecting ultra-low concentration analytes due to the inherent sensitivity limitations of LFIA.Recently,significant advances have been achieved by integrating enzyme activity probes and transforming LFIA into a highly sensitive tool for rapidly detecting trace analyte concentrations.Specifically,modifying natural enzymes or engineered nanozymes allows them to function as immune probes,directly catalyzing the production of signal molecules or indirectly initiating enzyme activity.Therefore,the signal intensity and detection sensitivity of LFIA are markedly elevated.The present review undertakes a comprehensive examination of pertinent research literature,offering a systematic analysis of recently proposed enzyme-based signal amplification strategies.By way of comparative assessment,the merits and demerits of current approaches are delineated,along with the identification of research avenues that still need to be explored.It is anticipated that this critical overview will garner considerable attention within the biomedical and materials science communities,providing valuable direction and insight toward the advancement of high-performance LFIA technologies.
基金supported by the National Natural Science Foundation of China(Nos.22064014,21765013)the Science and Technology Development Plan Project of Lanzhou(No.20211-146)+3 种基金the Science and Technology Project of Gansu Province(Nos.21YF5FA071,21JR7RA538)the Industrial Support Programme for Higher Education Institutions Project(Nos.2023CYZC-69,2024CYCZ-05)the 2023 Gansu Provincial Key Talent Project(No.2023RCXM26)a Gansu province postdoctoral grant(No.00247)。
文摘The advancement of various types of fluorescent nanoparticles is crucial for enhancing the application of lateral flow immunoassays(LFIA)across multiple fields.Currently,the fluorescent nanoparticles utilized in LFIA predominantly consist of traditional dye-doped nanoparticles or aggregation-induced luminescence dye-doped nanoparticles.The reliance on specific types of nanoparticles limits the diversity of signal reporting groups available for LFIA.Herein,we developed a solid-state luminescent dye-doped nanoparticles(SLDNPs)-based LFIA system with exceptional stability for the detection of C-reactive protein(CRP)in serum.The synthesis of SLD_(520)NP_(S)was simplicity,efficient and eco-friendly,which was ideal for large-scale production of the LFIA test strip.And the SLD_(520)NP_(S)exhibits superior fluorescence quantum yield(49%),fully guarantees the performance of the LFIA test strip.The constructed SLD_(520)NPsm Ab1-based LFIA demonstrated a satisfactory linear relationship with CRP concentrations ranging from 0.5 ng/mL to 100 ng/mL,with limits of detection(LOD)of 0.78 ng/mL and a visible LOD of 1 ng/mL using a handheld 405 nm lamp.Furthermore,the developed LFIA exhibited excellent recoveries in serum,ranging from 94.45%to 102.5%.Overall,the outstanding performance of the SLD_(520)NPs-mAb1-based LFIA indicates that solid-state luminescent dyes have significant potential applications in the field of LFIA.
基金financial support from the National Natural Science Foundation of China(Nos.22274022 and 21874022).
文摘Development of accurate analytical protocols for cancer biomarkers is used for the initial prescreening of malignant tumors,disease surveillance,and efficacy assessment with significant clinical benefits.In this work,we reported a liposome-mediated signal-off photoelectrochemical(PEC)immunoassay for the sensitive detection of carcinoembryonic antigen(CEA)using ternary transition metal sulfide CuS/ZnCdS as the photoactive material.Good photocurrents were acquired on the basis of specific oxidation reaction of dopamine on the CuS/ZnCdS.The energy band relationship of CuS/ZnCdS was determined,and the wellmatched oxidation potential of dopamine was verified.To achieve accurate recovery of low-abundance CEA,systematic PEC evaluation from human serum samples was performed by combining with classical immunoreaction and liposome-induced dopamine amplification strategy with high stability and selectivity.Under optimum conditions,PEC immunoassay displayed good photocurrent responses toward target CEA with a dynamic linear range of 0.1-50 ng/mL with a detection limit of 31.6 pg/mL.Importantly,this system by combining with a discussion of energy level matching between semiconductor energy bands and small-molecules opens a new horizon for development of high-efficient PEC immunoassays.
基金The Science and Technology Fund of Beijing Shijitan Hospital(Grant No.2022-C06)the National Key Research and Development Program of China(Grant No.2020YFF01014606).
文摘This study aimed to establish a highly accurate method for quantifying methotrexate(MTX)concentrations in serum using an ultra-high performance liquid chromatography-tandem mass spectrometry system(UPLC-MS/MS)and to compare its performance with the chemiluminescence microparticle immunoassay(CMIA).A total of 211 serum samples from pediatric patients with intracranial tumors undergoing high-dose MTX treatment were analyzed using both techniques.Correlation and agreement analyses were performed to assess the level of concordance between these methods.The results demonstrated a significant correlation(P<0.05)between the two methods,with correlation coefficients of 0.9913 and 0.9893,respectively.However,a statistical difference was noted in MTX serum concentrations at lower levels(0.04-1.5μmol/L),while no significant difference was observed at higher concentrations(1.5-400μmol/L).Specifically,in the 0.04-1.5μmol/L range(107 cases),Bland-Altman analysis indicated a bias of 0.018 between the two methods.Given the observed discrepancies,particularly at lower concentrations,it is advised that the UPLC-MS/MS method should not be used interchangeably with the CMIA assay for therapeutic drug monitoring in patients receiving high-dose MTX treatment.
基金funded by the Department of Biotechnology,Ministry of Science and Technology,Government of India(DBT)under grant number ADMaC DBT-NER/LIVS/11/2012.
文摘We report the development of a triplex nucleic acid lateral flow immunoassay(NALFIA)for the detection of the genomes of Nipah virus(NiV),Middle East respiratory syndrome coronavirus(MERS-CoV)and Reston ebolavirus(REBOV),which are intended for screening bats as well as other hosts and reservoirs of these three viruses.Our triplex NALFIA is a two-step assay format:the target nucleic acid in the sample is first amplified using tagged primers,and the tagged dsDNA amplicons are captured by antibodies immobilized on the NALFIA device,resulting in signal development from the binding of a streptavidin-colloidal gold conjugate to a biotin tag on the captured amplicons.Triplex amplification of the N gene of NiV,the UpE gene of MERS-CoV,and the Vp40 gene of REBOV was optimized,and three compatible combinations of hapten labels and antibodies were identified for end point detection.The lowest RNA copy numbers detected by the triplex NALFIA were 8.21e4 for the NiV N target,7.09e1 for the MERS-CoV UpE target,and 1.83e4 for the REBOV Vp40 target.Using simulated samples,the sensitivity and specificity for MERS-CoV and REBOV targets were estimated to be 100%,while the sensitivity and specificity for the NiV target were 91%and 93.3%,respectively.The compliance rate between triplex NALFIA and real-time RT‒PCR was 92%for the NiV N target and 100%for the MERS-CoV UpE and REBOV Vp40 targets.
基金supported by grants from Shanghai Agriculture Applied Technology Development Program,China(Grant No.:2020-02-08-00-08-F01456)the Key Research and Development Program of Zhejiang Province,China(Grant No.:2020C02024-2).
文摘The application of pesticides (mostly insecticides and fungicides) during the tea-planting process will undoubtedly increase the dietary risk associated with drinking tea. Thus, it is necessary to ascertain whether pesticide residues in tea products exceed the maximum residue limits. However, the complex matrices present in tea samples comprise a major challenge in the analytical detection of pesticide residues. In this study, nine types of lateral flow immunochromatographic strips (LFICSs) were developed to detect the pesticides of interest (fenpropathrin, chlorpyrifos, imidacloprid, thiamethoxam, acetamiprid, carbendazim, chlorothalonil, pyraclostrobin, and iprodione). To reduce the interference of tea substrates on the assay sensitivity, the pretreatment conditions for tea samples, including the extraction solvent, extraction time, and purification agent, were optimized for the simultaneous detection of these pesticides. The entire testing procedure (including pretreatment and detection) could be completed within 30 min. The detected results of authentic tea samples were confirmed by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), which suggest that the LFICS coupled with sample rapid pretreatment can be used for on-site rapid screening of the target pesticide in tea products prior to their market release.
基金supported financially by“Kunlun Talents High-end Innovation and Entrepreneurship Talents”of Qinghai Province in 2022National Natural Science Foundation of China(Nos.22322401 and 82073816)Beijing Nova Program(No.20220484055)。
文摘Foods are often contaminated by multiple foodborne pathogens,which threatens human health.In this work,we developed a microfluidic biosensor for multiplex immunoassay of foodborne bacteria with agitation driven by programmed audio signals.This agitation,powered by the vibration of a speaker cone during music playing,accelerated the mass transport in the incubation process to form bacterial complexes within 10 min.Immunoassay reagents of the two target bacteria(Escherichia coli O157:H7 and Salmonella typhimurium)were preloaded into the corresponding fore-vacuum storage chamber on the chip,and released to participate in the subsequent immune analysis process by piercing the chambers.All the detection processes were integrated into a single microfluidic chip and controlled by a smartphone through Bluetooth.Under selected conditions,wide linear ranges and low limits of detection(LODs<2CFU/m L)were obtained,and real food samples were successfully determined within 30 min.This biosensing method can be extended to wide-ranging applications by loading different recognizing reagents.
基金Supported by National Key R&D Program for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39)+2 种基金Key Research and Development Program of Shandong Province(Major Science and Technology Innovation Project)(2021CXGC011306)Scientific Research Project of General Administration of Customs(2024HK033)Scientific Research Project of Jinan Customs(2023JK005).
文摘[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.
文摘目的对人甲胎蛋白(alpha-fetoprotein,AFP)第2代国际标准品(编号:22/216)的含量进行协作标定研究。方法中国食品药品检定研究院代表中国实验室,组织了13家AFP试剂生产企业,采用13种化学发光免疫分析法、2种时间分辨荧光免疫法和1种酶联免疫法的试剂盒,按照英国药品和健康产品管理局提供的统一研究方案,以AFP第1代国际标准品为标准标定22/216。结果中国实验室提交的22/216含量的几何平均值(geometric mean,GM)为7828.78IU/安瓿(95%CI:7462.49~8213.05IU/安瓿),其几何变异系数(geometric coefficient of variation,GCV)为9.77%(n=16)。全球4个国家9个实验室参加了本研究,全部数据得到的22/216含量GM为7756.21IU/安瓿(95%CI:7486.81~8035.30IU/安瓿),GCV为8.73%(n=24)。结论经世界卫生组织生物标准专家委员会审核,确定22/216可作为AFP第2代国际标准品使用,效价为7800IU/安瓿。
