Glugea plecoglossi,a microsporidia of the Glugea genus,can cause an infamous disease Plecoglossus altivelis in East Asia,resulting in heavy economic losses.At present,the main diagnostic methods for this disease inclu...Glugea plecoglossi,a microsporidia of the Glugea genus,can cause an infamous disease Plecoglossus altivelis in East Asia,resulting in heavy economic losses.At present,the main diagnostic methods for this disease include microscopy examination,quantitative real-time PCR,and loop-mediated isothermal amplification-lateral flow dipstick(LAMP-LFD).In this study,a recombinase polymerase amplification-lateral flow dipstick(RPA-LFD)method,targeting the beta-tubulin gene,was developed to detect G.plecoglossi,three sets of primers and probes were designed and screened,after which the initial reaction system was established.The RPA-LFD method for G.plecoglossi could complete nucleic acid amplification at 39℃ for 10 min,after which the amplification product was dropped on the LFD strip,and the results could then be observed within 5 min.A specificity assay revealed that there was no cross reactivity with other protozoa except G.plecoglossi.A sensitivity assay revealed that the detection limit was 9.38×10^(-6) ng/μL,which was more sensitive than that of conventional PCR.Compared with conventional detection methods,the novel RPA-LFD method has the advantages of simple operation,short operation time,high sensitivity,and high specificity for G.plecoglossi detection,indicating its potential use in rapid field detection of G.plecoglossi.展开更多
BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited...BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited due to the organism’s fastidious growth requirements and prolonged culture time.AIM To propose a novel detection method utilizing antibiotic-supplemented media to inhibit susceptible strains,while resistant isolates were identified through urease-mediated hydrolysis of urea,inducing a phenol red color change for visual confirmation.METHODS Colombia agar was supplemented with urea,phenol red,and nickel chloride,and the final pH was adjusted to 7.35.Antibiotic-selective media were prepared by incorporating amoxicillin(0.5μg/mL),clarithromycin(2μg/mL),metronidazole(8μg/mL),or levofloxacin(2μg/mL)into separate batches.Gastric antral biopsies were homogenized and inoculated at 1.0×105 CFU onto the media,and then incubated under microaerobic conditions at 37°C for 28-36 hours.Resistance was determined based on a color change from yellow to pink,and the results were validated via broth microdilution according to Clinical and Laboratory Standards Institute guidelines.RESULTS After 28-36 hours of incubation,the drug-resistant H.pylori isolates induced a light red color change in the medium.Conversely,susceptible strains(H.pylori 26695 and G27)produced no visible color change.Compared with the conventional 11-day protocol,the novel method significantly reduced detection time.Among 201 clinical isolates,182 were successfully evaluated using the new method,resulting in a 90.5%detection rate.This was consistent with the 95.5%agreement rate observed when compared with microdilution-based susceptibility testing.The success rate of the novel approach was significantly higher than that of the comparative method(P<0.01).The accuracy of the new method was comparable to that of the dilution method.CONCLUSION The novel detection method can rapidly detect H.pylori drug resistance within 28-36 hours.With its operational simplicity and high diagnostic performance,it holds strong potential for clinical application in the management of H.pylori antimicrobial resistance.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
The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency ...The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency and insufficient sensitivity.This article reviews the progress of rapid detection technology for aquatic pathogens based on molecular biology,immunology,biosensors,etc.,and analyzes the application value of innovative methods such as isothermal amplification and CRISPR.This technology injects core momentum into the new quality productivity of fisheries through early and precise identification of pathogens:reducing aquaculture losses to improve resource efficiency,promoting the transformation of aquaculture models to data-driven,ensuring the safety of aquatic products to enhance the competitiveness of the industry chain.The current technology has shortcomings such as lagging standardization,weak on-site anti-interference ability,and insufficient recognition of new pathogens.In the future,we need to focus on technological integration and innovation,intelligent upgrading,and standardization construction,promote technology from laboratories to industrial applications,and provide continuous support for the high-quality development of fisheries.展开更多
Maize chlorotic dwarf virus （MCDV） is a quarantine pest as approved by Chinese government. A rapid, sensitive and specific MCDV detection method using reverse transcription-loop-mediated isothermal amplification （R...Maize chlorotic dwarf virus （MCDV） is a quarantine pest as approved by Chinese government. A rapid, sensitive and specific MCDV detection method using reverse transcription-loop-mediated isothermal amplification （RT-LAMP） was estab- lished in this study. Based on the sequence of MCDV coat protein coding gene, specific primers were designed and similar sensitivities were observed between RT- LAMP and RT-PCR, except that RT-LAMP was quicker, and the reaction could be finished within 1 h. In addition, the presence or absence of the fluorescent display in daylight allows naked easy detection of the amplification of MCDV genomic RNA using calcein. The RT-LAMP assay was applied successfully to detect MCDV in maize seeds, and the result by the addition of calcein was consistent with the result detected by the real time turbidimeter.展开更多
To relieve the increasing traffic load, many early built highways need to be widened or reconstructed. The rapid performance detection to existing subgrades is important to their reasonable evaluation and maximized ut...To relieve the increasing traffic load, many early built highways need to be widened or reconstructed. The rapid performance detection to existing subgrades is important to their reasonable evaluation and maximized utilization. Based on five kinds of soils taken from an existing highway in southern China, three commonly detecting methods were used to determine their moisture contents, compaction degrees and resilient moduli. The results showed that the measured moisture contents were greater than the design value, and the compaction degrees decreased sharply compared to the original ones. The moisture and heat exchange produced a decrease in the resilient modulus of plate loading test(PLT) from the standard 60 MPa down to 40 MPa. Afterwards, the portable falling weight deflectometer(PFWD) and dynamic cone penetrometer(DCP) were used to evaluate the subgrade performances. The measured PFWD moduli and the DCP penetration rates were correlated with the resilient moduli of PLT, deflections of the Beckman beam test, compaction degrees and moisture contents. The correlation analysis indicates that both of two methods are suitable in rapid detecting subgrade performances, but PFWD method is more recommended for it has higher accuracy and efficiency.展开更多
Chinese herbal medicines(CHMs)play an increasingly important role in the field of medicine and affects public health in the world.Although more and more strict has been employed to ensure the quality and safety of CHM...Chinese herbal medicines(CHMs)play an increasingly important role in the field of medicine and affects public health in the world.Although more and more strict has been employed to ensure the quality and safety of CHMs,pesticide residues in CHMs remain a serious issue and are the bottleneck for the global development of CHMs.In this work,we applied molecularly imprinted membrane electrospray mass spectrometry(MIM-ESI MS)for rapid detecting 4 classes of pesticide residues in CHMs,including organophosphorus(OPP),carbamates,pyrethroids and neonicotinoids in CHMs.Compared with our previous ambient ionization method MESI,MIM-ESI is capable of achieving a~50-fold increase in the detection limit of conventional analytical methods owing to the specificity recognition and unique enrichment of MIM.The optimal experimental conditions were determined,and the method was further validated for its sensitivity and specificity.Our data showed that MIM-ESI MS is applicable for the direct quantitation of pesticide residues in CHMs.This detection technology may help to ensure the quality of CHMs in the future.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
A novel electrochemical method for the rapid detection of organophosphorus pesticide residues was realized on a dual-channel screen-printed electrode （DSPE） that was integrated with a portable smartphone-controlled ...A novel electrochemical method for the rapid detection of organophosphorus pesticide residues was realized on a dual-channel screen-printed electrode （DSPE） that was integrated with a portable smartphone-controlled potentiostat. The two carbon working channels of DSPE were first modified by electrodepositing of Prussian blue. The channels were then modified with acetylcholinesterase （ACHE） via Nation. The inhibition ratio of AChE was detected by comparing the electrical current of acetylthiocholine （ATCh） that was catalyzed by the enzyme electrodes with （channel 1） and without （channel 2） organophosphorus pesticide. Inhibition ratios were related with the negative logarithm of the organophosphorus pesticide （trichlorfon, oxamyl, and isocarbophos） concentrations at optimum experimental conditions （pH 6.9 of electrolyte, 0.2V working potential, 2.5μL AChE modification amount, and 15 min inhibition time）. The linear equations were 1%=32.301gC＋ 253.3 （R=0.9750） for isocarbophos, I% = 35.991gC＋ 270.1 （R = 0.9668） for chlorpyrifos, and 1% = 33.701gC＋ 250.5 （R = 0.9606） for trichlorfon. The detection limits were calculated as 10-7 g/mL. Given that the inhibition ratios were only related with pesticide concentration and not with pesticide species, the proposed electrodes and electrometer can rapidly detect universal organophosphorus pesticides and assess pesticide pollution.展开更多
Pseudomonas aeruginosa(PA)is an opportunistic pathogen of humans and animals and a common source of nosocomial infections especially of the respiratory tract.Pseudomonas aeruginosa is also a major bacterial disease of...Pseudomonas aeruginosa(PA)is an opportunistic pathogen of humans and animals and a common source of nosocomial infections especially of the respiratory tract.Pseudomonas aeruginosa is also a major bacterial disease of poultry and in particular,eggs and newly hatched chicks.In this study,we developed a simple,accurate and rapid molecular detection method using cross priming amplification(CPA)with a nucleic acid test strip to detect P.aeruginosa.The assay efficiently amplified the target gene within 45 min at 62℃only using a simple water bath.The detection limit of the method was 1.18x 102 copiesμL^-1 for plasmid DNA and 4.4 CFU mL^-1 for bacteria in pure culture,and was 100 times more sensitive than conventional PCR.We screened 83 clinical samples from yellow-feather broiler breeder chickens and hospitalized/treated dogs and cats using CPA,PCR and traditional culture methods.The positive sample ratios were 15.3%(13/83)by CPA,13.3%(11/83)by PCR and 12.1%(10/83)by the culture method.The established CPA method has significant advantages for detecting P.aeruginosa.The method is easy to use and possesses high specificity and sensitivity without the requirements of complicated experimental equipment.The PA-CPA assay is especially fit for outdoor and primary medical units and is an ideal system for the rapid detection and monitoring of P.aeruginosa.展开更多
Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stran...Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides. Systematic evolution of ligands by exponential enrichment （SELEX） was used to select and PCR amplify DNA sequences （aptamers） capable of binding to and detecting Listeria monocytogenes, one of the major food-borne pathogens. A simplified affinity separation approach was employed, in which L. monocytogenes in exponential （log） phase of growth was used as the separation target. A fluorescently-labeled aptamer assay scheme was devised for detecting L. monoeytogenes. This report described a novel approach to the detection of L. monocytogenes using DNA aptamers. Aptamers were developed by nine rounds of SELEX. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. One of aptamers named e01 with the highest affinity was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. This study has proved the principle that the whole-cell SELEX could be a promising technique to design aptamer-based molecular probes for dectection of pathogenic microorganisms without tedious isolation and purification of complex markers or targets.展开更多
Rapid on-site detection of pathogenic bacteria with high sensitivity and specificity is becoming an urgent need in public health assurance, medical diagnostics, environmental monitoring, and food safety fields. Despit...Rapid on-site detection of pathogenic bacteria with high sensitivity and specificity is becoming an urgent need in public health assurance, medical diagnostics, environmental monitoring, and food safety fields. Despite being reliable and widely used, the existing methods of bacteria detection are cumbersome and time-consuming, which is not conducive to field detection. Microfluidic lab-on-a-chip technology has provided a detective tool for various analytes, due to its miniaturization, portability and low reagent consumption. Within this progress report, advances in the bacteria detection using microfluidic biosensors were discussed. Typical methods for pathogenic bacteria capture, separation and detection were introduced respectively in the first part. Then key applications of microfluidic biosensor-based rapid bacteria detection were presented. Finally, we made a conclusion and discussed possible research prospects in aspects of microfluidic biosensors for rapid detection of pathogenic bacteria.展开更多
[Objectives ] The paper was to explore enzyme inhibition rate method for rapid detection of organophosphorus and carbamate pesticides in cowpea. [ Methods ] Acetylcholinesterase （ACHE） was added to cowpea extract, t...[Objectives ] The paper was to explore enzyme inhibition rate method for rapid detection of organophosphorus and carbamate pesticides in cowpea. [ Methods ] Acetylcholinesterase （ACHE） was added to cowpea extract, to determine the inhibition rate of extract against enzyme. The influences of different sampiing methods and sampling parts on detection results were compared. [ Results] The positive rate of standard sampling was 18.18% higher than that of non-stand- ard sampling, and the positive rate of samples collected from cowpea tail was 16.67% higher than that collected from other parts. [ Condmions] Enzyme inhibi- tion rate method is suitable for rapid detection of organophosphorus and carbamate pesticides in cowpea.展开更多
Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. I...Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid （CA）/0.2 mol/L disodium hydrogen phosphate （NAP） buffer solution （pH 5.0）, 22 mmol/L 3,3＇,5,5＇-tetramethylbenzidine （TMB）, 0.6 mmol/L hydrogen peroxide （H202）, and 0.5% Tween-20 or 0.3% cetyltri- methyl ammonium bromide （CTAB） were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation （CV） of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation （c=0.9776）. So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.展开更多
Although detergent additives for gasoline have been widely commercialized,their formulas are often kept confidential and there is still no standardized method for quickly detecting the main active ingredients and eval...Although detergent additives for gasoline have been widely commercialized,their formulas are often kept confidential and there is still no standardized method for quickly detecting the main active ingredients and evaluating their effectiveness,which makes their regulation difficult.An overview of the current state of the development and application of detergent additives for gasoline in China and other regions,as well as a review of the rapid detection and performance evaluation methods available for analyzing detergent additives are given herein.The review focuses on the convenience,cost,efficiency,and feasibility of on-site detection and the evaluation of various methods,and also looks into future research directions,such as detecting and evaluating detergent additives in ethanol gasoline and with advanced engine technologies.展开更多
With the development of environmental monitoring,it is urgent to establish NO_(2)sensor with good sensing performance.Compared with the traditional NO_(2)sensors made of metal oxides,NO_(2)sensors made of n-p heterost...With the development of environmental monitoring,it is urgent to establish NO_(2)sensor with good sensing performance.Compared with the traditional NO_(2)sensors made of metal oxides,NO_(2)sensors made of n-p heterostructure nanocomposites have good sensing performance in detection limit and operating temperature.ZnO nanoflake arrays with polyaniline film grown on the surface were prepared on ceramic tubes by hydrothermal and vapor diffusion method.The gas-phase diffusion method can control the heterostructure by adjusting the diffusion time.At room temperature(25℃),the construction of rich n-p heterogeneous interface enables the sensor prepared by the nanocomposite to respond to NO_(2),showing the sensing performance with the response value of 28.00 to10.00×10^(-6)NO_(2);the detection limit improved to0.01×10^(-6)and the recovery time of 18 s.In this work,the sensing mechanism of NO_(2)at heterogeneous interface is analyzed,which provides a promising material for the detection of low concentration NO_(2)at room temperature.展开更多
Based on the chemical properties of dithiocarbamate pesticides,a device for rapid detection was developed in the paper,and the experimental conditions were optimized. Dithiocarbamate residues in fruits were successful...Based on the chemical properties of dithiocarbamate pesticides,a device for rapid detection was developed in the paper,and the experimental conditions were optimized. Dithiocarbamate residues in fruits were successfully detected using molecular absorption spectro-photometry,and the recovery rate was over 80%.The rapid detection method was simple to operate with low cost,and was conducive to application in basic level and enterprise laboratories.展开更多
Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 1...Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).展开更多
Dengue virus infections are increasing worldwide generally and in Asia,Central and South America and Africa,particularly.It poses a serious threat to the children population.The rapid and accurate diagnostic systems a...Dengue virus infections are increasing worldwide generally and in Asia,Central and South America and Africa,particularly.It poses a serious threat to the children population.The rapid and accurate diagnostic systems are essentially required due to lack of effective vaccine against dengue virus and the progressive spread of the dengue virus infection.The recent progress in developing micro-and nano-fabrication techniques has led to low cost and scale down the biomedical point-of-care devices.Starting from the conventional and modern available methods for the diagnosis of dengue infection,this review examines several emerging rapid and point-of-care diagnostic devices that hold significant potential for the progress in smart diagnosis tools.The given review revealed that an effective vaccine is required urgently against all the dengue virus serotypes.However,the rapid detection methods of dengue virus help in early treatment and significantly reduce the dengue virus outbreak.展开更多
Somatic cell count detection is the daily work of dairy farms to monitor the health of cows.The feasibility of applying near-infrared spectroscopy to somatic cell count detection was researched in this paper.Milk samp...Somatic cell count detection is the daily work of dairy farms to monitor the health of cows.The feasibility of applying near-infrared spectroscopy to somatic cell count detection was researched in this paper.Milk samples with different somatic cell counts were collected and preprocessing methods were studied.Variable selection algorithm based on hybrid strategy and modelling method based on ensemble learning were explored for somatic cell count detection.Detection model was used to diagnose subclinical mastitis and the results showed that near-infrared spectroscopy could be a tool to realize rapid detection of somatic cell count in milk.展开更多
基金Supported by the Visiting and Training Foundation of Teachers in Ordinary Undergraduate Universities of Shandong Province,the Qingdao Agricultural University Doctoral Start-Up Fund(No.6631122030)the Advanced Talents Foundation of QAU(No.6651118016)+2 种基金the Fish Innovation Team of Shandong Agriculture Research System(No.SDAIT12-06)the Shandong Engineering Research Center for Prevention and Control of Aquatic Animal Disease,the“First Class Fishery Discipline”Program[(2020)3]of Shandong Provincethe Key R&D Program(Soft Science Project)of Shandong Province,China(No.2023 RKY 06004)。
文摘Glugea plecoglossi,a microsporidia of the Glugea genus,can cause an infamous disease Plecoglossus altivelis in East Asia,resulting in heavy economic losses.At present,the main diagnostic methods for this disease include microscopy examination,quantitative real-time PCR,and loop-mediated isothermal amplification-lateral flow dipstick(LAMP-LFD).In this study,a recombinase polymerase amplification-lateral flow dipstick(RPA-LFD)method,targeting the beta-tubulin gene,was developed to detect G.plecoglossi,three sets of primers and probes were designed and screened,after which the initial reaction system was established.The RPA-LFD method for G.plecoglossi could complete nucleic acid amplification at 39℃ for 10 min,after which the amplification product was dropped on the LFD strip,and the results could then be observed within 5 min.A specificity assay revealed that there was no cross reactivity with other protozoa except G.plecoglossi.A sensitivity assay revealed that the detection limit was 9.38×10^(-6) ng/μL,which was more sensitive than that of conventional PCR.Compared with conventional detection methods,the novel RPA-LFD method has the advantages of simple operation,short operation time,high sensitivity,and high specificity for G.plecoglossi detection,indicating its potential use in rapid field detection of G.plecoglossi.
基金Supported by the Guangxi Science and Technology Major Projects,No.AA23073012the National Natural Science Foundation of China,No.32360035 and No.32060018。
文摘BACKGROUND Helicobacter pylori(H.pylori),a globally prevalent pathogen,is exhibiting increasing rates of antimicrobial resistance.However,clinical implementation of pre-treatment susceptibility testing remains limited due to the organism’s fastidious growth requirements and prolonged culture time.AIM To propose a novel detection method utilizing antibiotic-supplemented media to inhibit susceptible strains,while resistant isolates were identified through urease-mediated hydrolysis of urea,inducing a phenol red color change for visual confirmation.METHODS Colombia agar was supplemented with urea,phenol red,and nickel chloride,and the final pH was adjusted to 7.35.Antibiotic-selective media were prepared by incorporating amoxicillin(0.5μg/mL),clarithromycin(2μg/mL),metronidazole(8μg/mL),or levofloxacin(2μg/mL)into separate batches.Gastric antral biopsies were homogenized and inoculated at 1.0×105 CFU onto the media,and then incubated under microaerobic conditions at 37°C for 28-36 hours.Resistance was determined based on a color change from yellow to pink,and the results were validated via broth microdilution according to Clinical and Laboratory Standards Institute guidelines.RESULTS After 28-36 hours of incubation,the drug-resistant H.pylori isolates induced a light red color change in the medium.Conversely,susceptible strains(H.pylori 26695 and G27)produced no visible color change.Compared with the conventional 11-day protocol,the novel method significantly reduced detection time.Among 201 clinical isolates,182 were successfully evaluated using the new method,resulting in a 90.5%detection rate.This was consistent with the 95.5%agreement rate observed when compared with microdilution-based susceptibility testing.The success rate of the novel approach was significantly higher than that of the comparative method(P<0.01).The accuracy of the new method was comparable to that of the dilution method.CONCLUSION The novel detection method can rapidly detect H.pylori drug resistance within 28-36 hours.With its operational simplicity and high diagnostic performance,it holds strong potential for clinical application in the management of H.pylori antimicrobial resistance.
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 National Modern Agricultural Industry Technology System(CARS-45-33)Innovation Team of Tianjin Freshwater Aquaculture Industry Technology System(ITTFRS2021000-002,ITTFRS2021000-001)+1 种基金Tianjin Science and Technology Plan Project(24KPHDRC00280,24ZYCGSN00250,23YDTPJC00420)the Open Fund Project of Key Laboratory of Ocean Observation Technology,MNR(No.2023klootA03).
文摘The sustainable development of aquaculture industry is deeply constrained by pathogens and diseases,and traditional detection methods are difficult to adapt to the needs of intensive aquaculture due to low efficiency and insufficient sensitivity.This article reviews the progress of rapid detection technology for aquatic pathogens based on molecular biology,immunology,biosensors,etc.,and analyzes the application value of innovative methods such as isothermal amplification and CRISPR.This technology injects core momentum into the new quality productivity of fisheries through early and precise identification of pathogens:reducing aquaculture losses to improve resource efficiency,promoting the transformation of aquaculture models to data-driven,ensuring the safety of aquatic products to enhance the competitiveness of the industry chain.The current technology has shortcomings such as lagging standardization,weak on-site anti-interference ability,and insufficient recognition of new pathogens.In the future,we need to focus on technological integration and innovation,intelligent upgrading,and standardization construction,promote technology from laboratories to industrial applications,and provide continuous support for the high-quality development of fisheries.
文摘Maize chlorotic dwarf virus （MCDV） is a quarantine pest as approved by Chinese government. A rapid, sensitive and specific MCDV detection method using reverse transcription-loop-mediated isothermal amplification （RT-LAMP） was estab- lished in this study. Based on the sequence of MCDV coat protein coding gene, specific primers were designed and similar sensitivities were observed between RT- LAMP and RT-PCR, except that RT-LAMP was quicker, and the reaction could be finished within 1 h. In addition, the presence or absence of the fluorescent display in daylight allows naked easy detection of the amplification of MCDV genomic RNA using calcein. The RT-LAMP assay was applied successfully to detect MCDV in maize seeds, and the result by the addition of calcein was consistent with the result detected by the real time turbidimeter.
基金Project(2017YFC0805307) supported by the National Key Research and Development Program of ChinaProjects(51878078, 51927814, 51911530215) supported by the National Natural Science Foundation of China+4 种基金Project(2018-025) supported by the Training Program for High-level Technical Personnel in Transportation Industry, ChinaProject (2018JJ1026) supported by the Excellent Youth Foundation of Natural Science Foundation of Hunan Province, ChinaProject(17A008) supported by the Key Project of Education Department of Hunan Province, ChinaProjects(kfj150103, kfj170104) supported by the Open Research Fund of State Engineering Laboratory of Highway Maintenance Technology, Changsha University of Science & Technology, ChinaProject(CX20190644) supported by the Postgraduate Scientific Research Innovation Project of Hunan Province, China。
文摘To relieve the increasing traffic load, many early built highways need to be widened or reconstructed. The rapid performance detection to existing subgrades is important to their reasonable evaluation and maximized utilization. Based on five kinds of soils taken from an existing highway in southern China, three commonly detecting methods were used to determine their moisture contents, compaction degrees and resilient moduli. The results showed that the measured moisture contents were greater than the design value, and the compaction degrees decreased sharply compared to the original ones. The moisture and heat exchange produced a decrease in the resilient modulus of plate loading test(PLT) from the standard 60 MPa down to 40 MPa. Afterwards, the portable falling weight deflectometer(PFWD) and dynamic cone penetrometer(DCP) were used to evaluate the subgrade performances. The measured PFWD moduli and the DCP penetration rates were correlated with the resilient moduli of PLT, deflections of the Beckman beam test, compaction degrees and moisture contents. The correlation analysis indicates that both of two methods are suitable in rapid detecting subgrade performances, but PFWD method is more recommended for it has higher accuracy and efficiency.
基金supported by the National Natural Science Foundation of China(No.82072247)to MZthe Young Scholar Project of Beijing University of Chinese Medicine(Nos.2019-JYBJS-017 and 2021-JYB-XJSJJ001)to SJ and MZ。
文摘Chinese herbal medicines(CHMs)play an increasingly important role in the field of medicine and affects public health in the world.Although more and more strict has been employed to ensure the quality and safety of CHMs,pesticide residues in CHMs remain a serious issue and are the bottleneck for the global development of CHMs.In this work,we applied molecularly imprinted membrane electrospray mass spectrometry(MIM-ESI MS)for rapid detecting 4 classes of pesticide residues in CHMs,including organophosphorus(OPP),carbamates,pyrethroids and neonicotinoids in CHMs.Compared with our previous ambient ionization method MESI,MIM-ESI is capable of achieving a~50-fold increase in the detection limit of conventional analytical methods owing to the specificity recognition and unique enrichment of MIM.The optimal experimental conditions were determined,and the method was further validated for its sensitivity and specificity.Our data showed that MIM-ESI MS is applicable for the direct quantitation of pesticide residues in CHMs.This detection technology may help to ensure the quality of CHMs in the future.
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financially supported by the Natural Science Foundation of Zhejiang Province(No.LQ17B050002)Analysis and Measurement Foundation of Zhejiang Province(No.2015C37068)
文摘A novel electrochemical method for the rapid detection of organophosphorus pesticide residues was realized on a dual-channel screen-printed electrode （DSPE） that was integrated with a portable smartphone-controlled potentiostat. The two carbon working channels of DSPE were first modified by electrodepositing of Prussian blue. The channels were then modified with acetylcholinesterase （ACHE） via Nation. The inhibition ratio of AChE was detected by comparing the electrical current of acetylthiocholine （ATCh） that was catalyzed by the enzyme electrodes with （channel 1） and without （channel 2） organophosphorus pesticide. Inhibition ratios were related with the negative logarithm of the organophosphorus pesticide （trichlorfon, oxamyl, and isocarbophos） concentrations at optimum experimental conditions （pH 6.9 of electrolyte, 0.2V working potential, 2.5μL AChE modification amount, and 15 min inhibition time）. The linear equations were 1%=32.301gC＋ 253.3 （R=0.9750） for isocarbophos, I% = 35.991gC＋ 270.1 （R = 0.9668） for chlorpyrifos, and 1% = 33.701gC＋ 250.5 （R = 0.9606） for trichlorfon. The detection limits were calculated as 10-7 g/mL. Given that the inhibition ratios were only related with pesticide concentration and not with pesticide species, the proposed electrodes and electrometer can rapidly detect universal organophosphorus pesticides and assess pesticide pollution.
基金Supported by the Guangdong Key S&T Program(2019B020217002)from the Department of Science and Technology of Guangdong Province,China,the Guangdong Poultry Industry Technology System,China(2019KJ128)the earmarked fund for China Agriculture Research System(CARS-41-G16).
文摘Pseudomonas aeruginosa(PA)is an opportunistic pathogen of humans and animals and a common source of nosocomial infections especially of the respiratory tract.Pseudomonas aeruginosa is also a major bacterial disease of poultry and in particular,eggs and newly hatched chicks.In this study,we developed a simple,accurate and rapid molecular detection method using cross priming amplification(CPA)with a nucleic acid test strip to detect P.aeruginosa.The assay efficiently amplified the target gene within 45 min at 62℃only using a simple water bath.The detection limit of the method was 1.18x 102 copiesμL^-1 for plasmid DNA and 4.4 CFU mL^-1 for bacteria in pure culture,and was 100 times more sensitive than conventional PCR.We screened 83 clinical samples from yellow-feather broiler breeder chickens and hospitalized/treated dogs and cats using CPA,PCR and traditional culture methods.The positive sample ratios were 15.3%(13/83)by CPA,13.3%(11/83)by PCR and 12.1%(10/83)by the culture method.The established CPA method has significant advantages for detecting P.aeruginosa.The method is easy to use and possesses high specificity and sensitivity without the requirements of complicated experimental equipment.The PA-CPA assay is especially fit for outdoor and primary medical units and is an ideal system for the rapid detection and monitoring of P.aeruginosa.
文摘Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides. Systematic evolution of ligands by exponential enrichment （SELEX） was used to select and PCR amplify DNA sequences （aptamers） capable of binding to and detecting Listeria monocytogenes, one of the major food-borne pathogens. A simplified affinity separation approach was employed, in which L. monocytogenes in exponential （log） phase of growth was used as the separation target. A fluorescently-labeled aptamer assay scheme was devised for detecting L. monoeytogenes. This report described a novel approach to the detection of L. monocytogenes using DNA aptamers. Aptamers were developed by nine rounds of SELEX. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. One of aptamers named e01 with the highest affinity was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. This study has proved the principle that the whole-cell SELEX could be a promising technique to design aptamer-based molecular probes for dectection of pathogenic microorganisms without tedious isolation and purification of complex markers or targets.
基金supported by Research and Development Program in Key Areas of Guangdong Province,China (No.2019B020209009)the National Natural Science Foundation of China (Nos. 21727814, 81872829 and 21621003)。
文摘Rapid on-site detection of pathogenic bacteria with high sensitivity and specificity is becoming an urgent need in public health assurance, medical diagnostics, environmental monitoring, and food safety fields. Despite being reliable and widely used, the existing methods of bacteria detection are cumbersome and time-consuming, which is not conducive to field detection. Microfluidic lab-on-a-chip technology has provided a detective tool for various analytes, due to its miniaturization, portability and low reagent consumption. Within this progress report, advances in the bacteria detection using microfluidic biosensors were discussed. Typical methods for pathogenic bacteria capture, separation and detection were introduced respectively in the first part. Then key applications of microfluidic biosensor-based rapid bacteria detection were presented. Finally, we made a conclusion and discussed possible research prospects in aspects of microfluidic biosensors for rapid detection of pathogenic bacteria.
文摘[Objectives ] The paper was to explore enzyme inhibition rate method for rapid detection of organophosphorus and carbamate pesticides in cowpea. [ Methods ] Acetylcholinesterase （ACHE） was added to cowpea extract, to determine the inhibition rate of extract against enzyme. The influences of different sampiing methods and sampling parts on detection results were compared. [ Results] The positive rate of standard sampling was 18.18% higher than that of non-stand- ard sampling, and the positive rate of samples collected from cowpea tail was 16.67% higher than that collected from other parts. [ Condmions] Enzyme inhibi- tion rate method is suitable for rapid detection of organophosphorus and carbamate pesticides in cowpea.
基金supported by the National "Twelfth Five-Year" Plan for Science and Technology Support Program of China(No.2013BAD18B06)the Open Research Fund for the Key Laboratory of Dairy Science of Northeast Agricultural University(No.KLDS201201)the Innovative Research Team of Higher Education of Heilongjiang Province(No.2010td11),China
文摘Traditional methods for detecting lactoperoxidase （LP） are complex and time-consuming, so a test strip was made based on the enzymatic reaction principle to enable quick and convenient detection of LP in raw milk. In this study 0.1 mol/L citric acid （CA）/0.2 mol/L disodium hydrogen phosphate （NAP） buffer solution （pH 5.0）, 22 mmol/L 3,3＇,5,5＇-tetramethylbenzidine （TMB）, 0.6 mmol/L hydrogen peroxide （H202）, and 0.5% Tween-20 or 0.3% cetyltri- methyl ammonium bromide （CTAB） were optimal for preparing a quick, sensitive, and accurate LP test strip. The coefficient of variation （CV） of the estimated LP concentrations ranged from 2.47% to 6.72% and the minimum LP concentration detected by the test strip was 1-2 mg/L. Estimates of active LP in sixteen raw milk samples obtained using the test strip or the TMB method showed a good correlation （c=0.9776）. So the test strip provides a quick, convenient, and accurate method for detecting the LP concentration of raw milk.
基金This work was supported by the SINOPEC Research Project(No.121052-2).
文摘Although detergent additives for gasoline have been widely commercialized,their formulas are often kept confidential and there is still no standardized method for quickly detecting the main active ingredients and evaluating their effectiveness,which makes their regulation difficult.An overview of the current state of the development and application of detergent additives for gasoline in China and other regions,as well as a review of the rapid detection and performance evaluation methods available for analyzing detergent additives are given herein.The review focuses on the convenience,cost,efficiency,and feasibility of on-site detection and the evaluation of various methods,and also looks into future research directions,such as detecting and evaluating detergent additives in ethanol gasoline and with advanced engine technologies.
基金financially supported by the National Natural Science Foundation of China(Nos.21771060,61271126)the International Science&Technology Cooperation Program of China(No.2016YFE0115100)+1 种基金the Program for Science and Technology Project of Heilongjiang province(No.JQ2021B002)the Reform and Development Fund Project of Local University supported by the Central Government,Heilongjiang Touyan Innovation Team Program。
文摘With the development of environmental monitoring,it is urgent to establish NO_(2)sensor with good sensing performance.Compared with the traditional NO_(2)sensors made of metal oxides,NO_(2)sensors made of n-p heterostructure nanocomposites have good sensing performance in detection limit and operating temperature.ZnO nanoflake arrays with polyaniline film grown on the surface were prepared on ceramic tubes by hydrothermal and vapor diffusion method.The gas-phase diffusion method can control the heterostructure by adjusting the diffusion time.At room temperature(25℃),the construction of rich n-p heterogeneous interface enables the sensor prepared by the nanocomposite to respond to NO_(2),showing the sensing performance with the response value of 28.00 to10.00×10^(-6)NO_(2);the detection limit improved to0.01×10^(-6)and the recovery time of 18 s.In this work,the sensing mechanism of NO_(2)at heterogeneous interface is analyzed,which provides a promising material for the detection of low concentration NO_(2)at room temperature.
基金Supported by Class-A Projects of Fujian Department of Education(JA12465)Science and Technology Program of Xiamen City(3502Z20123046)
文摘Based on the chemical properties of dithiocarbamate pesticides,a device for rapid detection was developed in the paper,and the experimental conditions were optimized. Dithiocarbamate residues in fruits were successfully detected using molecular absorption spectro-photometry,and the recovery rate was over 80%.The rapid detection method was simple to operate with low cost,and was conducive to application in basic level and enterprise laboratories.
基金partially supported by the National Institutes of Health(grant no.P20GM103646)the United States Department of Agriculture Animal and Plant Health Inspection Service(agreement 14-7428-1041-CA)
文摘Dear Editor,Influenza A viruses(IAVs)are single-stranded,negative sense RNA viruses.IAV subtype is determined on the basis of the viral surface glycoproteins,hemagglutinin(HA),and neuraminidase(NA).To date,18 HA and 11NA subtypes have been reported(Tong et al.,2012).
基金supported by the Scientific Research Fund of the Shenzhen International cooperation projects under Grant Nos.(GJHZ20190819151403615)the Natural Science Youth Foundation of China(61801307).
文摘Dengue virus infections are increasing worldwide generally and in Asia,Central and South America and Africa,particularly.It poses a serious threat to the children population.The rapid and accurate diagnostic systems are essentially required due to lack of effective vaccine against dengue virus and the progressive spread of the dengue virus infection.The recent progress in developing micro-and nano-fabrication techniques has led to low cost and scale down the biomedical point-of-care devices.Starting from the conventional and modern available methods for the diagnosis of dengue infection,this review examines several emerging rapid and point-of-care diagnostic devices that hold significant potential for the progress in smart diagnosis tools.The given review revealed that an effective vaccine is required urgently against all the dengue virus serotypes.However,the rapid detection methods of dengue virus help in early treatment and significantly reduce the dengue virus outbreak.
基金Supported by the Natural Science Foundation of Heilongjiang Province of China(LH2023C016)the Key Research and Development Program of Heilongjiang Province of China(2022ZX01A24)the National Modern Agricultural Industry Technology System(CARS36)。
文摘Somatic cell count detection is the daily work of dairy farms to monitor the health of cows.The feasibility of applying near-infrared spectroscopy to somatic cell count detection was researched in this paper.Milk samples with different somatic cell counts were collected and preprocessing methods were studied.Variable selection algorithm based on hybrid strategy and modelling method based on ensemble learning were explored for somatic cell count detection.Detection model was used to diagnose subclinical mastitis and the results showed that near-infrared spectroscopy could be a tool to realize rapid detection of somatic cell count in milk.
