Fluorescence lateral flow immunoassay(LFA)has emerged as a powerful tool for rapid screening of various biomarkers owing to its simplicity,sensitivity and flexibility.It is noteworthy that fluorescent probe mainly det...Fluorescence lateral flow immunoassay(LFA)has emerged as a powerful tool for rapid screening of various biomarkers owing to its simplicity,sensitivity and flexibility.It is noteworthy that fluorescent probe mainly determines the analytical performance of LFA.Due to the emission and excitation wavelengths are located in the visible region,most fluorophores are inevitably subject to light scattering and background autofluorescence.Herein,we reported a novel LFA sensor based on the second near-infrared(NIR-Ⅱ)fluorescent probe with excellent anti-interference capability.The designed NIR-Ⅱprobe was the Nd^(3+)and Yb^(3+)doped rare earth nanoparticles(RENPs)by employing Nd^(3+)as energy donor and Yb^(3+)as energy acceptor,which of the donor-acceptor energy transfer(ET)efficiency reached up to 80.7%.Meanwhile,relying on the convenient and effective encapsulation strategy of poly(lactic-co-glycolic acid)(PLGA)microspheres to RENPs,the surface functionalized NIR-Ⅱprobe(RE@PLGA)was obtained for subsequent bioconjugation.Benefiting from the optical advantages of NIR-Ⅱprobe,this proposed NIR-ⅡLFA displayed a good linear relationship ranging from 7 ng/mL to 200 ng/mL for the detection ofα-fetoprotein(AFP),an important biomarker of hepatocellular carcinoma(HCC).The limit of detection(LOD)was determined as low as 3.0 ng/m L,which was of 8.3 times lower than clinical cutoff value.It is promising that LFA sensor based on this efficient RENPs probe provides new opportunities for high sensitive detection of various biomarkers in biological samples.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
Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The ext...Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The extensively used CSBV detection methods require professionals and expensive equipment;thus,they are unsuitable for rapid onsite CSBV detection.To achieve early and rapid detection of CSBV,we developed a lateral flow detection(LFD)strip method for CSBV detection via clustered regularly interspaced short palindromic repeats(CRISPR)and the Cas13a technique.On the basis of the conserved CSBV VP2 gene nucleotide region,we designed 3 recombinant enzyme-assisted amplification(RAA)primer pairs and prepared 3 corresponding crRNAs.We investigated key performance metrics,including the sensitivity,specificity,and accuracy of LFD strips.The results demonstrated that the LFD strip based on the optimal combination(primer 2+crRNA 2)presented the lowest detection limit(2.80×101 copies/μL),and this strip could complete CSBV detection within 1 h.Furthermore,this strip exhibited excellent detection specificity,with no cross-reactivity with four other honeybee viruses.A test of 100 clinical samples indicated the feasibility of the LFD method for CSBV detection.A comparison of various CSBV detection methods revealed that the CRISPR-Cas13a-based LFD method was more accurate,efficient,and sensitive than the other methods were,indicating great application prospects in onsite CSBV detection.Our developed method is highly important for preventing and controlling CSBV infection as well as maintaining honeybee health.展开更多
Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effective...Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effectiveness once BB has spread. Consequently, it is imperative to develop a rapid, highly sensitive, specific, and easy-to-use detection technique for early on-site diagnosis of BB. We first developed a recombinase-aided amplification-lateral flow dipstick(RAA-LFD) technique for the on-site detection of Xoo. The optimized reaction temperature and time were 37 ℃ and 20 min, indicating that the reaction system can be initiated by body temperature independently of any precision instruments. Evaluation of the RAA-LFD technique using the primers(RAAF2/R2) and probe(RAA2-nfo-probe) derived from the Xoo ORF0080 locus exhibited high specificity and eliminated cross-reactivity with other bacterial species. The sensitivity of RAA-LFD is up to 1 pg/μL for Xoo genomic DNA and 100 CFU/m L for Xoo cells. Significantly, this technique accurately detected Xoo from both artificially inoculated and naturally infected rice leaves at the early stage of infection, directly deploying plant tissue fluid as the template without DNA extraction. These attributes make the developed RAA-LFD system a viable technique for the early diagnosis of BB in the field, providing technical support for early-warning systems and disease control.展开更多
Porcine Contagious Pleuropneumonia(PCP)is a respiratory infectious disease of pigs caused by Actinobacillus pleuropneumoniae.The disease has been prevalent in pig farms since it was first identified in 1957(Pattison e...Porcine Contagious Pleuropneumonia(PCP)is a respiratory infectious disease of pigs caused by Actinobacillus pleuropneumoniae.The disease has been prevalent in pig farms since it was first identified in 1957(Pattison et al.1957).展开更多
Feline herpesvirus type 1(FHV-1)is a common and highly contagious pathogen in domestic cats that causes upper respiratory tract infections and ocular diseases.Accurate and rapid diagnosis of FHV-1 infections is essent...Feline herpesvirus type 1(FHV-1)is a common and highly contagious pathogen in domestic cats that causes upper respiratory tract infections and ocular diseases.Accurate and rapid diagnosis of FHV-1 infections is essential for effective disease management and control.In this study,we developed an immunochromatographic lateral flow(ICLF)assay for the rapid and accurate detection of FHV-1-specific antibodies.The assay was founded upon the successful expression and purification of a 26 kDa recombinant glycoprotein B-glycoprotein D(gB-gD)fusion protein,which served as the primary antigen for the test.Rigorous testing for specificity and cross-reactivity confirmed the strip’s ability to exclusively detect FHV-1 antibodies,even in the presence of a variety of other feline viruses.The assay demonstrated excellent precision,reproducibility across dilutions,and long-term stability,retaining efficacy for 24 months during storage.Furthermore,clinical sample analysis revealed exceptional sensitivity(97%)and specificity(100%).In conclusion,the ICLF strip developed in this study represents a reliable,highly specific,and stable diagnostic tool for the rapid detection and management of FHV-1 infections in cats.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
Lateral flow immunoassay(LFIA) is a widely used express method and offers advantages such as a short analysis time, simplicity of testing and result evaluation.However, an LFIA based on gold nanospheres lacks the desi...Lateral flow immunoassay(LFIA) is a widely used express method and offers advantages such as a short analysis time, simplicity of testing and result evaluation.However, an LFIA based on gold nanospheres lacks the desired sensitivity, thereby limiting its wide applications.In this study, spherical nanogold labels along with new types of nanogold labels such as gold nanopopcorns and nanostars were prepared, characterized, and applied for LFIA of model protein antigen procalcitonin. It was found that the label with a structure close to spherical provided more uniform distribution of specific antibodies on its surface, indicative of its suitability for this type of analysis.LFIA using gold nanopopcorns as a label allowed procalcitonin detection over a linear range of 0.5–10 ng mL^(-1) with the limit of detection of 0.1 ng mL^(-1), which was fivefold higher than the sensitivity of the assay with gold nanospheres. Another approach to improve the sensitivity of the assay included the silver enhancement method,which was used to compare the amplification of LFIA for procalcitonin detection. The sensitivity of procalcitonin determination by this method was 10 times better the sensitivity of the conventional LFIA with gold nanosphere as a label. The proposed approach of LFIA based on gold nanopopcorns improved the detection sensitivity without additional steps and prevented the increased consumption of specific reagents(antibodies).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
Lateral flow immunoassays(LFIAs)provide a powerful tool for rapid real-time assay of cancer biomarkers,which is vital for cancer detection and treatment follow-up.Lanthanide-based LFIAs is one of the most widely used ...Lateral flow immunoassays(LFIAs)provide a powerful tool for rapid real-time assay of cancer biomarkers,which is vital for cancer detection and treatment follow-up.Lanthanide-based LFIAs is one of the most widely used methods,especially Eu(Ⅲ)chelates,which possess distinctive and attractive characteristics,such as time-resolved fluorescence and large Stokes shift.Herein,we adopted a new onestep mini-emulsion polymerization method to synthesize carboxyl-modified fluorescent microsphere(OS-EuCM),which shows good stability,resistance to non-specific adhesion and uniform particle size distribution compared with traditional microspheres synthesized through the swelling method.Benefiting from the above advantages,OS-EuCM was successfully used in LFIAs to detect tumor marker Alpha-fetoprotein with high sensitivity and selectivity in concentration as high as 320 ng/mL,as well as a detection limit of 0.683 ng/mL This lanthanide-based microsphere holds great potential for rapid pointof-care screening and clinical application.展开更多
Recently, lateral flow assay （LFA） has attracted researchers＇ attention because of its considerable advantages of superior portability, rapid detection, cost-effectiveness and ease of use. This review provides a br...Recently, lateral flow assay （LFA） has attracted researchers＇ attention because of its considerable advantages of superior portability, rapid detection, cost-effectiveness and ease of use. This review provides a brief overview of latest researches of LFA, including the practical use of LFA in both qualitative and quantitative analysis in different areas. Though bio-recognition molecules in the LFA used to be antibodies, a new kind of recognition element called aptamer, showing significant advantages, is developed rapidly in recent years. The highly specific recognition of aptamers/antibodies and targets are combined with the excellent properties of a dry-reagent strip biosensor that enables efficiently detection in point-of-care applications. Herein, we compared the aptamers with antibodies, summarized the principle of LFAs, and three main elements for the LFAs （recognition molecule, signal transduction element, the targets）. Additionally, we summarized different optimal experimental conditions in the recent LFA-related studies to give detailed overview of the LFA development. We hope the review can give a general guide for the development of LFAs.展开更多
Lateral flow immunoassays(LFIAs) have been developed rapidly in recent years and used in a wide range of application at point-of-care-testing(POCT),where small biomolecules can be conveniently examined on a test strip...Lateral flow immunoassays(LFIAs) have been developed rapidly in recent years and used in a wide range of application at point-of-care-testing(POCT),where small biomolecules can be conveniently examined on a test strip.Compared with other biochemical detection methods such as ELISA(enzyme linked immunosorbent assay) or mass spectrometry method,LFIAs have the advantages of low cost,easy operation and short time-consuming.However,it suffers from low sensitivity since conventional LFIA can only realize qualitative detection based on colorimetric signals.With the increasing demand for more accurate and sensitive determination,novel nanomaterials have been used as labels in LFIAs due to their unique advantages in physical and chemical properties.Colloidal gold,fluorescent nano particles,SERSactive nanomaterials,magnetic nanoparticles and carbon nanomaterials are utilized in LFIAs to produce different kinds of signals for quantitative or semi-quantitative detection.This review paper first gives a description of the LFIA principles,and then focuses on the state-of-the-art nanomaterial labelling technology in LFIAs.At last,the conclusion and outlook are given to inspire exploration of more advanced nanomaterials for the development of future LFIAs.展开更多
A strip reader based lateral flow immunoassay （LFIA） was established for the rapid and quantitative detection of ractopamine （RAC） in swine urine. The ratio of the optical densities （ODs） of the test line （AT）...A strip reader based lateral flow immunoassay （LFIA） was established for the rapid and quantitative detection of ractopamine （RAC） in swine urine. The ratio of the optical densities （ODs） of the test line （AT） to that of the control line （Ac） was used to effectively minimize interference among strips and sample variations. The linear range for the quantitative detection of RAC was 0.2 ng/mL to 3.5 ng/mL with a median inhibitory concentration （IC50） of 0.59＋0.06 ng/mL. The limit of detection （LOD） of the LFIA was 0.13 ng/mL. The intra-assay recovery rates were 92.97%, 97.25%, and 107.41%, whereas the inter-assay rates were 80.07%, 108.17%, and 93.7%, respectively.展开更多
Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined ...Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)detection.Quantum dot nanobeads(QBs)served as fluorescence reporters,providing good detection efficiency.The customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product capture.Detection was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)line.The linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test strips.ST specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and SD-LAMP.We detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis.展开更多
In order to enhance the flavor of chafing dish and increase the attraction of consumers,the poppy shell is reported to be illegally added to the condiments of chafing dish.In this research,a rapid,simple,and convenien...In order to enhance the flavor of chafing dish and increase the attraction of consumers,the poppy shell is reported to be illegally added to the condiments of chafing dish.In this research,a rapid,simple,and convenient method based on the classic immunochromatographic lateral flow strip(LFS)with gold nanoparticles(GNPs)labeling was developed for easy monitoring of morphine(MOP),an effective component of poppy shell.Under optimized conditions,this developed LFS can well realize the detection of target MOP in the condiments of chafing dish in less than 10 min without any complicated pre-treatments.The limit of detection(LOD)can be achieved as low as 0.1 ppb for standard MOP or the MOP spiked condiments of chafing dish.All these results of the research strongly demonstrate that this established LFS method can be successfully applied in practical rapid and accurate on-site screening of poppy shell in condiments of chafing dish.展开更多
Objective To establish a sensitive,simple and rapid detection method for African swine fever virus(ASFV)B646L gene.Methods A recombinase-aided amplification-lateral flow dipstick(RAA-LFD)assay was developed in this st...Objective To establish a sensitive,simple and rapid detection method for African swine fever virus(ASFV)B646L gene.Methods A recombinase-aided amplification-lateral flow dipstick(RAA-LFD)assay was developed in this study.Recombinase-aided amplification(RAA)is used to amplify template DNA,and lateral flow dipstick(LFD)is used to interpret the results after the amplification is completed.The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid.In addition,30 clinical samples were tested to evaluate the performance of the RAA assay.Results The RAA-LFD assay was completed within 15 min at 37°C,including 10 min for nucleic acid amplification and 5 minutes for LFD reading results.The detection limit of this assay was found to be 200 copies per reaction.And there was no cross-reactivity with other swine viruses.Conclusion A highly sensitive,specific,and simple RAA-LFD method was developed for the rapid detection of the ASFV.展开更多
In this work,a novel conjugate of ractopamine and bovine serum albumin(RAC–BSA)has been developed via the Mannich reaction,with a mole coupling ratio for RAC–BSA of 9:1.The proposed conjugation method provides a sim...In this work,a novel conjugate of ractopamine and bovine serum albumin(RAC–BSA)has been developed via the Mannich reaction,with a mole coupling ratio for RAC–BSA of 9:1.The proposed conjugation method provides a simple and one-step method with the use of fewer reagents compared with other conjugation methods for competitive immunoassays.RAC–BSA conjugation was used to fabricate a competitive lateral flow strip test for RAC detection in animal feed.For sample preparation,RAC was spiked in swine feed purchased from the local markets in Thailand,and methanol and running buffer at a volume ratio of 10:90 was used as extraction buffer.The procedures for sample preparation were completed within 25 min.Under optimal conditions,the limit of detection(LOD),assessed by the naked eye within 5 min,was found to be 1 ng/g.A semi-quantitative analysis was also conducted using a smart phone and computer software,with a linearity of 0.075–0.750 ng/g,calculated LOD of 0.10 ng/g,calculated limit of quantitation of 0.33 ng/g,and good correlation of 0.992.The recoveries were found in the range of 96.4%–103.7%with a relative standard deviation of 2.5%–3.6%for intra-and inter-assays.Comparison of the results obtained by the strip test with those obtained by enzyme-linked immunosorbent assay had a good agreement in terms of accuracy.Furthermore,this strip test exhibited highly specific RAC detection without cross reactivity with related compounds.Therefore,the RAC–BSA conjugation via the Mannich reaction can be accepted as a one-step and easy conjugation method and applied to the competitive lateral flow strip test.展开更多
基金supported by the National Natural Science Foundation of China(Nos.U2267221,22107029,22377135)the Bohai Rim Advanced Research Institute for Drug Discovery(No.LX215002)+5 种基金the Natural Science Foundation of Shandong Province(No.ZR2022QH212)the Taishan Scholars Program(No.tsqn202312305)the Young Elite Scientists Sponsorship Program by Chinese Chemical Societythe Fundamental Research Projects of Science&Technology Innovation and development Plan in Yantai City(No.2023JCYJ059)the Shandong Laboratory Program(No.SYS202205)the Shanghai Postdoctoral Excellence Program(No.2023704)。
文摘Fluorescence lateral flow immunoassay(LFA)has emerged as a powerful tool for rapid screening of various biomarkers owing to its simplicity,sensitivity and flexibility.It is noteworthy that fluorescent probe mainly determines the analytical performance of LFA.Due to the emission and excitation wavelengths are located in the visible region,most fluorophores are inevitably subject to light scattering and background autofluorescence.Herein,we reported a novel LFA sensor based on the second near-infrared(NIR-Ⅱ)fluorescent probe with excellent anti-interference capability.The designed NIR-Ⅱprobe was the Nd^(3+)and Yb^(3+)doped rare earth nanoparticles(RENPs)by employing Nd^(3+)as energy donor and Yb^(3+)as energy acceptor,which of the donor-acceptor energy transfer(ET)efficiency reached up to 80.7%.Meanwhile,relying on the convenient and effective encapsulation strategy of poly(lactic-co-glycolic acid)(PLGA)microspheres to RENPs,the surface functionalized NIR-Ⅱprobe(RE@PLGA)was obtained for subsequent bioconjugation.Benefiting from the optical advantages of NIR-Ⅱprobe,this proposed NIR-ⅡLFA displayed a good linear relationship ranging from 7 ng/mL to 200 ng/mL for the detection ofα-fetoprotein(AFP),an important biomarker of hepatocellular carcinoma(HCC).The limit of detection(LOD)was determined as low as 3.0 ng/m L,which was of 8.3 times lower than clinical cutoff value.It is promising that LFA sensor based on this efficient RENPs probe provides new opportunities for high sensitive detection of various biomarkers in biological samples.
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supported by the National Key Research and Development Program of China(NO.2022YFD1600202)the Lush Mountain Public Welfare Protection Action of the China Environmental Protection Foundation(CEPFQS202169-14).
文摘Sacbrood virus(SBV)is one of the most pathogenic honeybee viruses with host specificity and regional variation.The SBV strain infecting the Chinese honeybee(Apis cerana)is known as Chinese sacbrood virus(CSBV).The extensively used CSBV detection methods require professionals and expensive equipment;thus,they are unsuitable for rapid onsite CSBV detection.To achieve early and rapid detection of CSBV,we developed a lateral flow detection(LFD)strip method for CSBV detection via clustered regularly interspaced short palindromic repeats(CRISPR)and the Cas13a technique.On the basis of the conserved CSBV VP2 gene nucleotide region,we designed 3 recombinant enzyme-assisted amplification(RAA)primer pairs and prepared 3 corresponding crRNAs.We investigated key performance metrics,including the sensitivity,specificity,and accuracy of LFD strips.The results demonstrated that the LFD strip based on the optimal combination(primer 2+crRNA 2)presented the lowest detection limit(2.80×101 copies/μL),and this strip could complete CSBV detection within 1 h.Furthermore,this strip exhibited excellent detection specificity,with no cross-reactivity with four other honeybee viruses.A test of 100 clinical samples indicated the feasibility of the LFD method for CSBV detection.A comparison of various CSBV detection methods revealed that the CRISPR-Cas13a-based LFD method was more accurate,efficient,and sensitive than the other methods were,indicating great application prospects in onsite CSBV detection.Our developed method is highly important for preventing and controlling CSBV infection as well as maintaining honeybee health.
基金supported by the Zhejiang Provincial Natural Science Foundation of China(Grant Nos.LY23C130004 and LZ24C130004)the National Natural Science Foundation of China(Grant No.32472115)+1 种基金the National Key Research and Development Program of China(Grant No.2022YFF1003301)the Agricultural Sciences and Technologies Innovation Program of Chinese Academy of Agricultural Sciences。
文摘Bacterial blight(BB) is a devastating worldwide rice disease caused by Xanthomonas oryzae pv. oryzae(Xoo), which is difficult to diagnose based on early symptoms. Conventional chemical control yields limited effectiveness once BB has spread. Consequently, it is imperative to develop a rapid, highly sensitive, specific, and easy-to-use detection technique for early on-site diagnosis of BB. We first developed a recombinase-aided amplification-lateral flow dipstick(RAA-LFD) technique for the on-site detection of Xoo. The optimized reaction temperature and time were 37 ℃ and 20 min, indicating that the reaction system can be initiated by body temperature independently of any precision instruments. Evaluation of the RAA-LFD technique using the primers(RAAF2/R2) and probe(RAA2-nfo-probe) derived from the Xoo ORF0080 locus exhibited high specificity and eliminated cross-reactivity with other bacterial species. The sensitivity of RAA-LFD is up to 1 pg/μL for Xoo genomic DNA and 100 CFU/m L for Xoo cells. Significantly, this technique accurately detected Xoo from both artificially inoculated and naturally infected rice leaves at the early stage of infection, directly deploying plant tissue fluid as the template without DNA extraction. These attributes make the developed RAA-LFD system a viable technique for the early diagnosis of BB in the field, providing technical support for early-warning systems and disease control.
基金supported by the University-Industry Col aborative Education Program,China(220904860093831)。
文摘Porcine Contagious Pleuropneumonia(PCP)is a respiratory infectious disease of pigs caused by Actinobacillus pleuropneumoniae.The disease has been prevalent in pig farms since it was first identified in 1957(Pattison et al.1957).
基金supported by the Fundamental Research Program of Shanxi Province (202403021221077)the Central Funds Guiding the Local Science and Technology Development in Shanxi Province (YDZJSX2021 A034)+3 种基金the Project of Scientific Research for Excellent Doctors,Shanxi Province,China(SXBYKY2021047)the Research Fund (Clinical Diagnosis and Treatment of Pet) for Young College Teachers in Ruipeng Commonwealth Foundation (RPJJ2020021)the Project of Science and Technology Innovation Fund of Shanxi Agricultural University (2021BQ06)the Inner Mongolia Autonomous Region First Class Discipline Research Special Project(YLXKZX-NND-012)
文摘Feline herpesvirus type 1(FHV-1)is a common and highly contagious pathogen in domestic cats that causes upper respiratory tract infections and ocular diseases.Accurate and rapid diagnosis of FHV-1 infections is essential for effective disease management and control.In this study,we developed an immunochromatographic lateral flow(ICLF)assay for the rapid and accurate detection of FHV-1-specific antibodies.The assay was founded upon the successful expression and purification of a 26 kDa recombinant glycoprotein B-glycoprotein D(gB-gD)fusion protein,which served as the primary antigen for the test.Rigorous testing for specificity and cross-reactivity confirmed the strip’s ability to exclusively detect FHV-1 antibodies,even in the presence of a variety of other feline viruses.The assay demonstrated excellent precision,reproducibility across dilutions,and long-term stability,retaining efficacy for 24 months during storage.Furthermore,clinical sample analysis revealed exceptional sensitivity(97%)and specificity(100%).In conclusion,the ICLF strip developed in this study represents a reliable,highly specific,and stable diagnostic tool for the rapid detection and management of FHV-1 infections in cats.
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financially supported by the Ministry of Education and Science of the Russian Federation in the framework of increase Competitiveness Program of NUST ‘‘MISIS’’, implemented by a governmental decree dated 16th of March 2013, No. 211part of state assignment Organization of scientific researches (Project No. 16.6548.2017/BY)
文摘Lateral flow immunoassay(LFIA) is a widely used express method and offers advantages such as a short analysis time, simplicity of testing and result evaluation.However, an LFIA based on gold nanospheres lacks the desired sensitivity, thereby limiting its wide applications.In this study, spherical nanogold labels along with new types of nanogold labels such as gold nanopopcorns and nanostars were prepared, characterized, and applied for LFIA of model protein antigen procalcitonin. It was found that the label with a structure close to spherical provided more uniform distribution of specific antibodies on its surface, indicative of its suitability for this type of analysis.LFIA using gold nanopopcorns as a label allowed procalcitonin detection over a linear range of 0.5–10 ng mL^(-1) with the limit of detection of 0.1 ng mL^(-1), which was fivefold higher than the sensitivity of the assay with gold nanospheres. Another approach to improve the sensitivity of the assay included the silver enhancement method,which was used to compare the amplification of LFIA for procalcitonin detection. The sensitivity of procalcitonin determination by this method was 10 times better the sensitivity of the conventional LFIA with gold nanosphere as a label. The proposed approach of LFIA based on gold nanopopcorns improved the detection sensitivity without additional steps and prevented the increased consumption of specific reagents(antibodies).
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Project supported by the National Key R&D Program of China(2017YFA0205100)。
文摘Lateral flow immunoassays(LFIAs)provide a powerful tool for rapid real-time assay of cancer biomarkers,which is vital for cancer detection and treatment follow-up.Lanthanide-based LFIAs is one of the most widely used methods,especially Eu(Ⅲ)chelates,which possess distinctive and attractive characteristics,such as time-resolved fluorescence and large Stokes shift.Herein,we adopted a new onestep mini-emulsion polymerization method to synthesize carboxyl-modified fluorescent microsphere(OS-EuCM),which shows good stability,resistance to non-specific adhesion and uniform particle size distribution compared with traditional microspheres synthesized through the swelling method.Benefiting from the above advantages,OS-EuCM was successfully used in LFIAs to detect tumor marker Alpha-fetoprotein with high sensitivity and selectivity in concentration as high as 320 ng/mL,as well as a detection limit of 0.683 ng/mL This lanthanide-based microsphere holds great potential for rapid pointof-care screening and clinical application.
基金financially support to the Key Program of Beijing Municipal Science and Technology Commission (No. D161100002116002)the National Key Research and Development Program of China (Nos. 2016YFF0203703, 2017YFB0306901)the National Natural Science Foundation of China (No. 51673012)
文摘Recently, lateral flow assay （LFA） has attracted researchers＇ attention because of its considerable advantages of superior portability, rapid detection, cost-effectiveness and ease of use. This review provides a brief overview of latest researches of LFA, including the practical use of LFA in both qualitative and quantitative analysis in different areas. Though bio-recognition molecules in the LFA used to be antibodies, a new kind of recognition element called aptamer, showing significant advantages, is developed rapidly in recent years. The highly specific recognition of aptamers/antibodies and targets are combined with the excellent properties of a dry-reagent strip biosensor that enables efficiently detection in point-of-care applications. Herein, we compared the aptamers with antibodies, summarized the principle of LFAs, and three main elements for the LFAs （recognition molecule, signal transduction element, the targets）. Additionally, we summarized different optimal experimental conditions in the recent LFA-related studies to give detailed overview of the LFA development. We hope the review can give a general guide for the development of LFAs.
基金financial support from the National Natural Science Foundation of China(51802060)Shenzhen Science and Technology Program(Grant No.:KQTD20170809110344233)+1 种基金Shenzhen Bay Laboratory(SZBL2019062801005)Natural Science Foundation of Guangdong Province(No.2019A1515010762)。
文摘Lateral flow immunoassays(LFIAs) have been developed rapidly in recent years and used in a wide range of application at point-of-care-testing(POCT),where small biomolecules can be conveniently examined on a test strip.Compared with other biochemical detection methods such as ELISA(enzyme linked immunosorbent assay) or mass spectrometry method,LFIAs have the advantages of low cost,easy operation and short time-consuming.However,it suffers from low sensitivity since conventional LFIA can only realize qualitative detection based on colorimetric signals.With the increasing demand for more accurate and sensitive determination,novel nanomaterials have been used as labels in LFIAs due to their unique advantages in physical and chemical properties.Colloidal gold,fluorescent nano particles,SERSactive nanomaterials,magnetic nanoparticles and carbon nanomaterials are utilized in LFIAs to produce different kinds of signals for quantitative or semi-quantitative detection.This review paper first gives a description of the LFIA principles,and then focuses on the state-of-the-art nanomaterial labelling technology in LFIAs.At last,the conclusion and outlook are given to inspire exploration of more advanced nanomaterials for the development of future LFIAs.
基金supported by the national science and technology support program in the 12th Five Year Plan(2011BAK10B04 and 2011BAK10B01)the national natural science foundation of China(Grant No.31160323)the research program of the state key laboratory of food science and technology,Nanchang University(SKLF-ZZB-201306)
文摘A strip reader based lateral flow immunoassay （LFIA） was established for the rapid and quantitative detection of ractopamine （RAC） in swine urine. The ratio of the optical densities （ODs） of the test line （AT） to that of the control line （Ac） was used to effectively minimize interference among strips and sample variations. The linear range for the quantitative detection of RAC was 0.2 ng/mL to 3.5 ng/mL with a median inhibitory concentration （IC50） of 0.59＋0.06 ng/mL. The limit of detection （LOD） of the LFIA was 0.13 ng/mL. The intra-assay recovery rates were 92.97%, 97.25%, and 107.41%, whereas the inter-assay rates were 80.07%, 108.17%, and 93.7%, respectively.
基金This work was supported by the National Key Research and Development Program of China(2019YFC1606300)the Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(2017BT01S174)the Guangdong Academy of Sciences Special Project of Implementing Innovation-Driven Development Capacity Building(2018GDASCX-0401).
文摘Rapid,sensitive,point-of-care detection of pathogenic bacteria is important for food safety.In this study,we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip(QBs-labelled LFIAS)combined with strand displacement loop-mediated isothermal amplification(SD-LAMP)for quantitative Salmonella Typhimurium(ST)detection.Quantum dot nanobeads(QBs)served as fluorescence reporters,providing good detection efficiency.The customizable strand displacement(SD)probe was used in LAMP to improve the specificity of the method and prevent by-product capture.Detection was based on a sandwich immunoassay.A fluorescence strip reader measured the fluorescence intensity(FI)of the test(T)line and control(C)line.The linear detection range of the strip was 10^(2)–10^(8) colony forming units(CFU)·mL^(-1).The visual limit of detection was 10^(3) CFU·mL^(-1),indicating that the system was ten-fold more sensitive than AuNPs-labelled test strips.ST specificity was analyzed in accordance with agarose gel outputs of polymerase chain reaction(PCR)and SD-LAMP.We detected ST in foods with an acceptable recovery of 85%–110%.The method is rapid,simple,almost equipment-free,and suitable for bacterial detection in foods and for clinical diagnosis.
基金the National Natural Science Foundation of China(Grants No.21475030,21804028)the Fundamental Research Fund for central university(Grants No.2017HGPA0162,JZ2018HGTA0205,PA2017GDQT0018)+2 种基金the grant of 2017YFF0208600,the China Agriculture Research System-48(CARS-48)Anhui Provincial Modern Argo-industry Tech.Research System(NYCYTX-2016-84)the S&T Research Project of Anhui Province(Grant No.15czz03109).
文摘In order to enhance the flavor of chafing dish and increase the attraction of consumers,the poppy shell is reported to be illegally added to the condiments of chafing dish.In this research,a rapid,simple,and convenient method based on the classic immunochromatographic lateral flow strip(LFS)with gold nanoparticles(GNPs)labeling was developed for easy monitoring of morphine(MOP),an effective component of poppy shell.Under optimized conditions,this developed LFS can well realize the detection of target MOP in the condiments of chafing dish in less than 10 min without any complicated pre-treatments.The limit of detection(LOD)can be achieved as low as 0.1 ppb for standard MOP or the MOP spiked condiments of chafing dish.All these results of the research strongly demonstrate that this established LFS method can be successfully applied in practical rapid and accurate on-site screening of poppy shell in condiments of chafing dish.
基金supported by National Key R&D Program of China[2017YFC200503]National Natural Science Foundation of China[No.42077399].
文摘Objective To establish a sensitive,simple and rapid detection method for African swine fever virus(ASFV)B646L gene.Methods A recombinase-aided amplification-lateral flow dipstick(RAA-LFD)assay was developed in this study.Recombinase-aided amplification(RAA)is used to amplify template DNA,and lateral flow dipstick(LFD)is used to interpret the results after the amplification is completed.The lower limits of detection and specificity of the RAA assay were verified using recombinant plasmid and pathogenic nucleic acid.In addition,30 clinical samples were tested to evaluate the performance of the RAA assay.Results The RAA-LFD assay was completed within 15 min at 37°C,including 10 min for nucleic acid amplification and 5 minutes for LFD reading results.The detection limit of this assay was found to be 200 copies per reaction.And there was no cross-reactivity with other swine viruses.Conclusion A highly sensitive,specific,and simple RAA-LFD method was developed for the rapid detection of the ASFV.
基金Project Project supported by the Thailand Research Fund through the International Research Network(No.PHD58W0001)the Thailand Research Fund through Research Team Promotion Grant the Ratchadaphisaksomphot Endowment Fund of Chulalongkorn University(No.RTA6080002),Thailand
文摘In this work,a novel conjugate of ractopamine and bovine serum albumin(RAC–BSA)has been developed via the Mannich reaction,with a mole coupling ratio for RAC–BSA of 9:1.The proposed conjugation method provides a simple and one-step method with the use of fewer reagents compared with other conjugation methods for competitive immunoassays.RAC–BSA conjugation was used to fabricate a competitive lateral flow strip test for RAC detection in animal feed.For sample preparation,RAC was spiked in swine feed purchased from the local markets in Thailand,and methanol and running buffer at a volume ratio of 10:90 was used as extraction buffer.The procedures for sample preparation were completed within 25 min.Under optimal conditions,the limit of detection(LOD),assessed by the naked eye within 5 min,was found to be 1 ng/g.A semi-quantitative analysis was also conducted using a smart phone and computer software,with a linearity of 0.075–0.750 ng/g,calculated LOD of 0.10 ng/g,calculated limit of quantitation of 0.33 ng/g,and good correlation of 0.992.The recoveries were found in the range of 96.4%–103.7%with a relative standard deviation of 2.5%–3.6%for intra-and inter-assays.Comparison of the results obtained by the strip test with those obtained by enzyme-linked immunosorbent assay had a good agreement in terms of accuracy.Furthermore,this strip test exhibited highly specific RAC detection without cross reactivity with related compounds.Therefore,the RAC–BSA conjugation via the Mannich reaction can be accepted as a one-step and easy conjugation method and applied to the competitive lateral flow strip test.
