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
Rapid detection of target foodborne pathogens plays more and more significant roles in food safety,which requires the efficiency,sensitivity,and accuracy.In this research,we proposed a new st rategy of isothermal-mole...Rapid detection of target foodborne pathogens plays more and more significant roles in food safety,which requires the efficiency,sensitivity,and accuracy.In this research,we proposed a new st rategy of isothermal-molecular-amplification integrated with lateral-flow-strip for rapid detection of Salmonella without traditional enrichment-culture.Th e designed syringe-assisted-filtration can contribute to simultaneous collection and concentration of target bacterium from vegetable samples in just 3 min,resolving the drawbacks of traditional random sampling protocols.After simple and convenient ultrasonication,samples can be directly amplified at 39℃ in 25 min and the amplicons are qualitatively and quantitatively analyzed with the designed lateral-flow-strip in 5 min.Finally,satisfied results have been achieved within 40 min,which greatly improve the efficiency while the accuracy is also guaranteed.Furthermore,all detection steps can be completed under instrument-free conditions.This method will hold great promise for target pathogen detection in the resource-limited district,or for emergency on-site identification.展开更多
Ureaplasma urealyticum(UU),is one of the most vital pathogens causing genitourinary tract infections of the body,and it can result in poor maternal and perinatal outcomes.The aim of this study was to establish a metho...Ureaplasma urealyticum(UU),is one of the most vital pathogens causing genitourinary tract infections of the body,and it can result in poor maternal and perinatal outcomes.The aim of this study was to establish a method to detect Ureaplasma urealyticum based on recombinant polymerase amplification(RPA)technique.Specific primers and probes were designed according to the 16sRNA gene sequence of Ureaplasma urealyticum.Six pathogens were detected for real-time fluorescence RPA specificity verification,including Mycoplasma hominis(MH),Chlamydia trachomatis(CT),Neisseria gonorrhoeae(NG),Staphylococcus aureus,Escherichia coli,and Lactobacillus vaginalis.The sensitivity of the method was performed by gradient dilution of the extracted template.A total of 60 clinical samples were detected by the established real-time fluorescence RPA.Detection of Ureaplasma urealyticum can be completed within 20 minutes at 39°C using established RPA method.The minimum detection limit of Ureaplasma urealyticum by real-time fluorescence RPA was 3 pg.The evaluation of 60 clinical samples proved that RPA method was feasible.A high specificity,sensitivity,simplicity and rapidity method for Ureaplasma urealyticum detection was successfully established based on the real-time fluorescence RPA method.展开更多
Objective:To evaluate luciferase reporter phage(LRP)phAE85 in rapid detection of rifampicin resistance in a region where TB is endemic.Methods:One hundred and ninety primary isolates on Lowenstein-Jensen medium were t...Objective:To evaluate luciferase reporter phage(LRP)phAE85 in rapid detection of rifampicin resistance in a region where TB is endemic.Methods:One hundred and ninety primary isolates on Lowenstein-Jensen medium were tested.Middlebrook 7H9 complete medium with and without rifampicin at 2μg/mL was inoculated with standard inoculum from suspensions of the clinical isolate.After incubation for 72 h,LRP was added.Following 4 h of further incubation,light output from both control and test was measured as relative light units.Strains exhibiting a reduction of less than 50%relative light units in the drug containing vial compared to control were classified as resistant.Results were compared with the conventional minimum inhibitory concentration method(MIC)of drug susceptibility testing.Results:The two methods showed high level of agreement of 97%(CI 0.94,0.99)and P value was 0.000 1.The sensitivity and specificity of LRP assay for detection of rifampicin resistance were 91%h(CI 0.75,0.98)and 99ct(CI0.95,1.00)respectively.Time to detection of resistance by LRP assay was 3 d in comparison with 28 d by the minimum inhibitory concentration method.Conclusions:LRP assay with phAE85 is 99%specific,91%sensitive and is highly reproducible.Thus the assay offers a simple procedure for drug sensitivity testing,within die scope of semi-automation.展开更多
[Objectives] This study was conducted to establish a rapid detection method for spectinomycin in pork,chicken,fish,shrimp flesh and water.[Methods]A test strip for rapid detection of spectinomycin in milk was develope...[Objectives] This study was conducted to establish a rapid detection method for spectinomycin in pork,chicken,fish,shrimp flesh and water.[Methods]A test strip for rapid detection of spectinomycin in milk was developed by colloidal gold immunochromatography assay. [Results]The test strip had a detection limit of 50 μg/kg to milk with a detection time of 15 min,and the false positive rate and false negative rate were both 0. [Conclusions]The method is accurate,simple,reliable and convenient,and is suitable for rapid on-site spectinomycin detection.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
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
Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the head...Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced,disease control by fungicide application solely based on the detection of visual symptoms is ineffective.To develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control,we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments,MoT-6098 and MoT-6099,that are present in the MoT genome but not in the genome of the rice-infecting Magnaporthe oryzae Oryzae(MoO)pathotype.Using polymerase chain reaction(PCR),we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and Bangladesh.To test the efficiency of the two markers,we first established a loop-mediated isothermal amplification(LAMP)method to detect MoT at isothermal conditions,without the use of a PCR machine.Following this,we used the Cas12a protein and guide RNAs(gRNAs)to target the MoT-6098 and MoT-6099 sequences.The activated Cas12a showed indiscriminate single-stranded deoxyribonuclease(ssDNase)activity.We then combined targetdependent Cas12a ssDNase activation with recombinase polymerase amplification(RPA)and nucleic acid lateral flow immunoassay(NALFIA)to develop a method that accurately,sensitively,and cost-effectively detects MoT-specific DNA sequences in infected wheat plants.This novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.展开更多
Exposure to toxins can lead to a wide range of adverse health effects, including respiratory problems, neurological disorders, cancer, and reproductive issues. Toxins can come from various sources, such as industrial ...Exposure to toxins can lead to a wide range of adverse health effects, including respiratory problems, neurological disorders, cancer, and reproductive issues. Toxins can come from various sources, such as industrial waste, agricultural runoff, and household chemicals. Therefore, detecting and monitoring toxins in the environment is crucial for protecting human health and the environment. This study aimed to evaluate the performance of Hememics biosensor system in detecting environmental toxins such as Ricin and Staphylococcal enterotoxin B (SEB) in mixed matrixes. When Ricin and SEB are spiked into soil, chopped lettuce, tap water, milk and serum, the biosensor was able to detect these toxins, without sample processing, at a level of detection comparable to lab testing with high sensitivity and specificity. Furthermore, Hememics biosensor system is designed to be network-enabled, which means that results can be transmitted to relevant agencies for quick decisions. This feature is crucial in cases where quick action is needed to prevent further contamination or exposure to harmful toxins.展开更多
Earthquake detection and location are essential in earthquake studies,which generally consists of two main classes:waveform-based and pick-based methods.To evaluate the ability of two different methods,a graphicsproce...Earthquake detection and location are essential in earthquake studies,which generally consists of two main classes:waveform-based and pick-based methods.To evaluate the ability of two different methods,a graphicsprocessing-unit-based Match&Locate(GPU-M&L)method and a rapid earthquake association and location(REAL)method are applied to continuous seismic data recorded by 24 digital seismic stations from Jiangsu Seismic Network during 2013 for comparison.GPU-M&L is one of waveform-based methods by waveform cross-correlations while REAL is one of pick-based method to associate arrivals of different seismic phases and locate events through counting the number of P and S picks and travel time residuals.Twenty-six templates are selected from the Jiangsu Seismic Network local catalog by using the GPU-M&L.The number of newly detected and located events is about 2.8 times more than those listed in the local catalog.We both utilize a deep-neural-network-based arrival-time picking method called PhaseNet and a shortterm/long-term average(STA/LTA)trigger algorithm for seismic phase detection and picking by applying the REAL.We then refine seismic locations using a least-squares location method(VELEST)and a high-precision relative location method(hypoDD).By applying STA/LTA and PhaseNet,1006 and 1893 events are associated and located,respectively.The newly detected events are mainly clustered and show steeply dipping fault planes.By analyzing the performance of these methods based on long-term continuous seismic data,the detected catalogs by the GPU-M&L and REAL show that the magnitudes of completeness are 1.4 and 0.8,respectively,which are smaller than 2.6 given by the local catalog.Although REAL provides improvement compared with GPU-M&L,REAL is highly dependent on phase detection and picking which is strongly affected by signal-noise ratio(SNR).Stations at southeast of the study region with low SNR may lead to few detections in the same area.展开更多
A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the ...A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV. PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.展开更多
Onlineγ-spectrometry systems for inland waters,most of which extract samples in situ and in real time,are able to produce reliable activity concentration measurements for waterborne radionuclides only when they are d...Onlineγ-spectrometry systems for inland waters,most of which extract samples in situ and in real time,are able to produce reliable activity concentration measurements for waterborne radionuclides only when they are distributed relatively uniformly and enter into a steady-state diffusion regime in the measurement chamber.To protect residents’health and ensure the safety of the living environment,better timeliness is required for this measurement method.To address this issue,this study established a mathematical model of the online waterγ-spectrometry system so that rapid warning and activity estimates can be obtained for water under non-steady-state(NSS)conditions.In addition,the detection efficiency of the detector for radionuclides during the NSS diffusion process was determined by applying the computational fluid dynamics technique in conjunction with Monte Carlo simulations.On this basis,a method was developed that allowed the online waterγ-spectrometry system to provide rapid warning and activity concentration estimates for radionuclides in water.Subsequent analysis of the NSS-mode measurements of^(40)K radioactive solutions with different activity concentrations determined the optimum warning threshold and measurement time for producing accurate activity concentration estimates for radionuclides.The experimental results show that the proposed NSS measurement method is able to give warning and yield accurate activity concentration estimates for radionuclides 55.42 and 69.42 min after the entry of a 10 Bq/L^(40)K radioactive solution into the measurement chamber,respectively.These times are much shorter than the 90 min required by the conventional measurement method.Furthermore,the NSS measurement method allows the measurement system to give rapid(within approximately 15 min)warning when the activity concentrations of some radionuclides reach their respective limits stipulated in the Guidelines for Drinking-water Quality of the WHO,suggesting that this method considerably enhances the warning capacity of in situ online waterγ-spectrometry systems.展开更多
Objective:To analyse molecular detection of coliforms and shorten the time of PCR.Methods:Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in ...Objective:To analyse molecular detection of coliforms and shorten the time of PCR.Methods:Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number(MPN)method for 16 artificial and 101 field samples.The molecular method was also conducted on isolated coliforms from positive MPN samples;standard sample for verification of microbial method certificated reference material;isolated strains from certificated reference material and standard bacteria.The PCR and electrophoresis parameters were changed for reducing the operation time.Results:Results of PCR for lacZ and uidA genes were similar in all of standard,operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR.PCR results were confirmed by MPN culture method by sensitivity 86%(95%CI:0.71-0.93).Also the total execution time,with a successful change of factors,was reduced to less than two and a half hour.Conclusions:Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city.It's recommended to be used at least as an initial screening test,and then the positive samples could be randomly tested by MPN.展开更多
A novel electrochemical immunoassay for cardiac troponin Ⅰ (cTnI) combining the concepts of the dual monoclonal antibody "sandwich" principle, the silver enhancement on the nano-gold particle, and the SBA-15 meso...A novel electrochemical immunoassay for cardiac troponin Ⅰ (cTnI) combining the concepts of the dual monoclonal antibody "sandwich" principle, the silver enhancement on the nano-gold particle, and the SBA-15 mesoporous modified carbon paste electrode (SBA-MCPE) is described. Four main steps were carried out to obtain the analytical signal, i.e., electrode preparation, immunoreaction, silver enhancement, and anodic stripping voltammetric detection. A linear relationship between the anodic stripping peak current and concentration of cTnI from 0.5 to 5.0 ng/mL and a limit of detection of 0.2 ng/mL of cTnI were obtained.展开更多
For the first time, mass spectrometric (MS) techniques were employed to rapidly detect the pathogen Chalara fraxinea in-vitro and directly in-vivo in tissues of diseased ash trees caused by C. fraxinea, using a range ...For the first time, mass spectrometric (MS) techniques were employed to rapidly detect the pathogen Chalara fraxinea in-vitro and directly in-vivo in tissues of diseased ash trees caused by C. fraxinea, using a range of characteristic novel secondary metabolites of C. fraxinea as chemical markers for the presence of the pathogen. We have found an evident correlation between the presence and amount of these-only for C. fraxinea characteristic and novel-secondary metabolites (named chalarafraxinines) and the degree of disease of respective infected ash seedlings. As demonstrated in this work, the MS based high-throughput-screening approach constitute an alternative to the time consuming and expensive micro biological isolation procedures for detection of the pathogen C. fraxinea and furthermore, can be used to rapidly test ash genotypes for resistance / susceptibility to C. fraxinea infection.展开更多
Meloidogyne vitis is a new root-knot nematode parasitic on grape root in Yunnan Province,China.In order to establish a rapid,reliable and specific molecular detection method for M.vitis,the species-specific primers we...Meloidogyne vitis is a new root-knot nematode parasitic on grape root in Yunnan Province,China.In order to establish a rapid,reliable and specific molecular detection method for M.vitis,the species-specific primers were designed with rDNA-ITS(ribosomal DNA internal transcribed spacer)gene fragment as the target.The reaction system was optimized and the reliability,specificity and sensitivity of primer were testified,therefore,a rapid PCR detection method for M.vitis was established.The result showed that the optimal annealing temperature of the primers was 53℃,which was suitable for the detection of different life stages of M.vitis.Specificity test showed that the specific fragment size of 174 bp was obtained from M.vitis,but other five non-target nematodes did not have any amplification bands,thus effectively distinguish M.vitis and the other five species,and could specifically detect the M.vitis from mixed populations.Sensitivity test showed that this PCR technique could detect the DNA of a single second-stage juvenile(J_(2))and 10^(-4)female.Futhermore,this PCR technique could be used to detect directly M.vitis from soil samples.The rapid,sensitive and specific PCR molecular detection technique could be used for the direct identification of a single J_(2)of M.vitis and the detection of M.vitis in mixed nematode populations and the detection of two J_(2)s or one male in 0.5 g soil samples,which will provide technical support for the investigation of the occurrence and damage of M.vitis and the formulation of efficient green co ntrol strategies.展开更多
Enteroaggregative Escherichia coli (EAggEC) strains cause the persistent diarrhea in infants and compromised hosts in developing countries. These strains are currently defined as E. coli that adheres to HEp-2 cells in...Enteroaggregative Escherichia coli (EAggEC) strains cause the persistent diarrhea in infants and compromised hosts in developing countries. These strains are currently defined as E. coli that adheres to HEp-2 cells in an aggregative adherence (AA) pattern. In this study, we compared 4 different rapid methods for the detection of EAggEC using a PCR assay, clump formation test, glass slide adherence assay, and the HEp-2 cell adherence assay. Out of 683 E. coli strains isolated from diarrheal stool samples, we detected 17 aggR and/or clump-positive strains, and identified 2 aggR-positive, clump-negative strains and 2 aggR-negative, clump-positive strains. All the aggR positive and clump positive strains also showed positive results in glass slide adherence and HEp-2 cell adherence assays. From all these results, we suggest the following procedure for the rapid identification of EAggEC strains: first, screen E. coli strains with the clump formation test and subsequently perform the glass slide adherence assay to observe AA for confirmation.展开更多
Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs...Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs), disease detection has become easier and faster. However, these RDTs have failed to compete with the specialized laboratory equipment due to their high detection limits and false alarm rates. This paper presents a novel method of using carbon nanofibers (CNFs) grown on glass microballoons (NMBs) to achieve ultra-low detection limits in RDTs. The NMBs have millions of nanosized CNFs grown on each microballoon, with each CNF having a strong bonding affinity for antibodies. The NMBs conjugated with secondary antibodies have therefore a significantly higher probability of capturing minute antigen concentrations in solution. Furthermore, the dark color formation at the capture zone makes visual disease detection possible. Human Immunoglobulin G (IgG) was selected as the model analyte to study the performance of NMBs using a sandwich immunoassay protocol. Ultra-low electrical detection limit of (4 pg/ml) and rapid re- sponse (~1 minute) was achieved using this method.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
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 grants of the NSFC(32172295,21804028)the key R&D program of Anhui(201904d07020016)+5 种基金the Anhui Provincial NSF(1908085QC121)the Fundamental Research Fund for central university(JZ2019HGTB0068)the China Postdoctoral Science Foundation(2019M652167)the Fund of State Key Lab of Chemo/Biosensing and Chemometrics(Hunan University),the postdoc grant of Anhui(2020B412)Young and Middle-aged Leading Scientists,Engineers and Innovators of the XPCC(2019CB017)China Agriculture Research System-48(CARS-48).
文摘Rapid detection of target foodborne pathogens plays more and more significant roles in food safety,which requires the efficiency,sensitivity,and accuracy.In this research,we proposed a new st rategy of isothermal-molecular-amplification integrated with lateral-flow-strip for rapid detection of Salmonella without traditional enrichment-culture.Th e designed syringe-assisted-filtration can contribute to simultaneous collection and concentration of target bacterium from vegetable samples in just 3 min,resolving the drawbacks of traditional random sampling protocols.After simple and convenient ultrasonication,samples can be directly amplified at 39℃ in 25 min and the amplicons are qualitatively and quantitatively analyzed with the designed lateral-flow-strip in 5 min.Finally,satisfied results have been achieved within 40 min,which greatly improve the efficiency while the accuracy is also guaranteed.Furthermore,all detection steps can be completed under instrument-free conditions.This method will hold great promise for target pathogen detection in the resource-limited district,or for emergency on-site identification.
文摘Ureaplasma urealyticum(UU),is one of the most vital pathogens causing genitourinary tract infections of the body,and it can result in poor maternal and perinatal outcomes.The aim of this study was to establish a method to detect Ureaplasma urealyticum based on recombinant polymerase amplification(RPA)technique.Specific primers and probes were designed according to the 16sRNA gene sequence of Ureaplasma urealyticum.Six pathogens were detected for real-time fluorescence RPA specificity verification,including Mycoplasma hominis(MH),Chlamydia trachomatis(CT),Neisseria gonorrhoeae(NG),Staphylococcus aureus,Escherichia coli,and Lactobacillus vaginalis.The sensitivity of the method was performed by gradient dilution of the extracted template.A total of 60 clinical samples were detected by the established real-time fluorescence RPA.Detection of Ureaplasma urealyticum can be completed within 20 minutes at 39°C using established RPA method.The minimum detection limit of Ureaplasma urealyticum by real-time fluorescence RPA was 3 pg.The evaluation of 60 clinical samples proved that RPA method was feasible.A high specificity,sensitivity,simplicity and rapidity method for Ureaplasma urealyticum detection was successfully established based on the real-time fluorescence RPA method.
文摘Objective:To evaluate luciferase reporter phage(LRP)phAE85 in rapid detection of rifampicin resistance in a region where TB is endemic.Methods:One hundred and ninety primary isolates on Lowenstein-Jensen medium were tested.Middlebrook 7H9 complete medium with and without rifampicin at 2μg/mL was inoculated with standard inoculum from suspensions of the clinical isolate.After incubation for 72 h,LRP was added.Following 4 h of further incubation,light output from both control and test was measured as relative light units.Strains exhibiting a reduction of less than 50%relative light units in the drug containing vial compared to control were classified as resistant.Results were compared with the conventional minimum inhibitory concentration method(MIC)of drug susceptibility testing.Results:The two methods showed high level of agreement of 97%(CI 0.94,0.99)and P value was 0.000 1.The sensitivity and specificity of LRP assay for detection of rifampicin resistance were 91%h(CI 0.75,0.98)and 99ct(CI0.95,1.00)respectively.Time to detection of resistance by LRP assay was 3 d in comparison with 28 d by the minimum inhibitory concentration method.Conclusions:LRP assay with phAE85 is 99%specific,91%sensitive and is highly reproducible.Thus the assay offers a simple procedure for drug sensitivity testing,within die scope of semi-automation.
基金Supported by Beijing Training Project for the Leading Talents in Science and Technology(Z171100001117158)
文摘[Objectives] This study was conducted to establish a rapid detection method for spectinomycin in pork,chicken,fish,shrimp flesh and water.[Methods]A test strip for rapid detection of spectinomycin in milk was developed by colloidal gold immunochromatography assay. [Results]The test strip had a detection limit of 50 μg/kg to milk with a detection time of 15 min,and the false positive rate and false negative rate were both 0. [Conclusions]The method is accurate,simple,reliable and convenient,and is suitable for rapid on-site spectinomycin detection.
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 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The data that support the findings of this study have being submitted to GenBank and the accession numbers are JAAXMV000000000 and JAAXMU000000000.
文摘Wheat blast,caused by the fungus Magnaporthe oryzae Triticum(MoT)pathotype,is a devastating disease persistent in South America and Bangladesh.Since MoT generally fails to cause visual symptoms in wheat until the heading stage when the infection would have advanced,disease control by fungicide application solely based on the detection of visual symptoms is ineffective.To develop an accurate and sensitive method to detect MoT at the seedling and vegetative stages for disease control,we sequenced the genomes of two MoT isolates from Brazil and identified two DNA fragments,MoT-6098 and MoT-6099,that are present in the MoT genome but not in the genome of the rice-infecting Magnaporthe oryzae Oryzae(MoO)pathotype.Using polymerase chain reaction(PCR),we confirmed the specificity of the two markers in 53 MoT and MoO isolates from South America and Bangladesh.To test the efficiency of the two markers,we first established a loop-mediated isothermal amplification(LAMP)method to detect MoT at isothermal conditions,without the use of a PCR machine.Following this,we used the Cas12a protein and guide RNAs(gRNAs)to target the MoT-6098 and MoT-6099 sequences.The activated Cas12a showed indiscriminate single-stranded deoxyribonuclease(ssDNase)activity.We then combined targetdependent Cas12a ssDNase activation with recombinase polymerase amplification(RPA)and nucleic acid lateral flow immunoassay(NALFIA)to develop a method that accurately,sensitively,and cost-effectively detects MoT-specific DNA sequences in infected wheat plants.This novel technique can be easily adapted for the rapid detection of wheat blast and other important plant diseases in the field.
文摘Exposure to toxins can lead to a wide range of adverse health effects, including respiratory problems, neurological disorders, cancer, and reproductive issues. Toxins can come from various sources, such as industrial waste, agricultural runoff, and household chemicals. Therefore, detecting and monitoring toxins in the environment is crucial for protecting human health and the environment. This study aimed to evaluate the performance of Hememics biosensor system in detecting environmental toxins such as Ricin and Staphylococcal enterotoxin B (SEB) in mixed matrixes. When Ricin and SEB are spiked into soil, chopped lettuce, tap water, milk and serum, the biosensor was able to detect these toxins, without sample processing, at a level of detection comparable to lab testing with high sensitivity and specificity. Furthermore, Hememics biosensor system is designed to be network-enabled, which means that results can be transmitted to relevant agencies for quick decisions. This feature is crucial in cases where quick action is needed to prevent further contamination or exposure to harmful toxins.
基金This research is co-supported by National Key R&D Program of China(No.2017YFC1500402)National Natural Science Foundation of China(Nos.41874063 and U1939203)Shanghai Sheshan National Geophysical Observatory(No.2020K02)。
文摘Earthquake detection and location are essential in earthquake studies,which generally consists of two main classes:waveform-based and pick-based methods.To evaluate the ability of two different methods,a graphicsprocessing-unit-based Match&Locate(GPU-M&L)method and a rapid earthquake association and location(REAL)method are applied to continuous seismic data recorded by 24 digital seismic stations from Jiangsu Seismic Network during 2013 for comparison.GPU-M&L is one of waveform-based methods by waveform cross-correlations while REAL is one of pick-based method to associate arrivals of different seismic phases and locate events through counting the number of P and S picks and travel time residuals.Twenty-six templates are selected from the Jiangsu Seismic Network local catalog by using the GPU-M&L.The number of newly detected and located events is about 2.8 times more than those listed in the local catalog.We both utilize a deep-neural-network-based arrival-time picking method called PhaseNet and a shortterm/long-term average(STA/LTA)trigger algorithm for seismic phase detection and picking by applying the REAL.We then refine seismic locations using a least-squares location method(VELEST)and a high-precision relative location method(hypoDD).By applying STA/LTA and PhaseNet,1006 and 1893 events are associated and located,respectively.The newly detected events are mainly clustered and show steeply dipping fault planes.By analyzing the performance of these methods based on long-term continuous seismic data,the detected catalogs by the GPU-M&L and REAL show that the magnitudes of completeness are 1.4 and 0.8,respectively,which are smaller than 2.6 given by the local catalog.Although REAL provides improvement compared with GPU-M&L,REAL is highly dependent on phase detection and picking which is strongly affected by signal-noise ratio(SNR).Stations at southeast of the study region with low SNR may lead to few detections in the same area.
基金The National Science and Technology supporting plan of the Eleventh Five-year(2006BAD06A18 and 2006BAD06A03)Beijing Natural Science Foundation(5072041)
文摘A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV. PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.
基金supported by the National Natural Science Foundation of China(No.42127807)Natural Science Foundation of Sichuan Province of China(Project No.2023NSFSC0008)+1 种基金Uranium Geology Program of China Nuclear Geology(No.202205-6)the Sichuan Science and Technology Program(No.2021JDTD0018)。
文摘Onlineγ-spectrometry systems for inland waters,most of which extract samples in situ and in real time,are able to produce reliable activity concentration measurements for waterborne radionuclides only when they are distributed relatively uniformly and enter into a steady-state diffusion regime in the measurement chamber.To protect residents’health and ensure the safety of the living environment,better timeliness is required for this measurement method.To address this issue,this study established a mathematical model of the online waterγ-spectrometry system so that rapid warning and activity estimates can be obtained for water under non-steady-state(NSS)conditions.In addition,the detection efficiency of the detector for radionuclides during the NSS diffusion process was determined by applying the computational fluid dynamics technique in conjunction with Monte Carlo simulations.On this basis,a method was developed that allowed the online waterγ-spectrometry system to provide rapid warning and activity concentration estimates for radionuclides in water.Subsequent analysis of the NSS-mode measurements of^(40)K radioactive solutions with different activity concentrations determined the optimum warning threshold and measurement time for producing accurate activity concentration estimates for radionuclides.The experimental results show that the proposed NSS measurement method is able to give warning and yield accurate activity concentration estimates for radionuclides 55.42 and 69.42 min after the entry of a 10 Bq/L^(40)K radioactive solution into the measurement chamber,respectively.These times are much shorter than the 90 min required by the conventional measurement method.Furthermore,the NSS measurement method allows the measurement system to give rapid(within approximately 15 min)warning when the activity concentrations of some radionuclides reach their respective limits stipulated in the Guidelines for Drinking-water Quality of the WHO,suggesting that this method considerably enhances the warning capacity of in situ online waterγ-spectrometry systems.
基金Supported by the Ministry of Power of I.R.Iran(Grant No.201)
文摘Objective:To analyse molecular detection of coliforms and shorten the time of PCR.Methods:Rapid detection of coliforms by amplification of lacZ and uidA genes in a multiplex PCR reaction was designed and performed in comparison with most probably number(MPN)method for 16 artificial and 101 field samples.The molecular method was also conducted on isolated coliforms from positive MPN samples;standard sample for verification of microbial method certificated reference material;isolated strains from certificated reference material and standard bacteria.The PCR and electrophoresis parameters were changed for reducing the operation time.Results:Results of PCR for lacZ and uidA genes were similar in all of standard,operational and artificial samples and showed the 876 bp and 147 bp bands of lacZ and uidA genes by multiplex PCR.PCR results were confirmed by MPN culture method by sensitivity 86%(95%CI:0.71-0.93).Also the total execution time,with a successful change of factors,was reduced to less than two and a half hour.Conclusions:Multiplex PCR method with shortened operation time was used for the simultaneous detection of total coliforms and Escherichia coli in distribution system of Arak city.It's recommended to be used at least as an initial screening test,and then the positive samples could be randomly tested by MPN.
文摘A novel electrochemical immunoassay for cardiac troponin Ⅰ (cTnI) combining the concepts of the dual monoclonal antibody "sandwich" principle, the silver enhancement on the nano-gold particle, and the SBA-15 mesoporous modified carbon paste electrode (SBA-MCPE) is described. Four main steps were carried out to obtain the analytical signal, i.e., electrode preparation, immunoreaction, silver enhancement, and anodic stripping voltammetric detection. A linear relationship between the anodic stripping peak current and concentration of cTnI from 0.5 to 5.0 ng/mL and a limit of detection of 0.2 ng/mL of cTnI were obtained.
文摘For the first time, mass spectrometric (MS) techniques were employed to rapidly detect the pathogen Chalara fraxinea in-vitro and directly in-vivo in tissues of diseased ash trees caused by C. fraxinea, using a range of characteristic novel secondary metabolites of C. fraxinea as chemical markers for the presence of the pathogen. We have found an evident correlation between the presence and amount of these-only for C. fraxinea characteristic and novel-secondary metabolites (named chalarafraxinines) and the degree of disease of respective infected ash seedlings. As demonstrated in this work, the MS based high-throughput-screening approach constitute an alternative to the time consuming and expensive micro biological isolation procedures for detection of the pathogen C. fraxinea and furthermore, can be used to rapidly test ash genotypes for resistance / susceptibility to C. fraxinea infection.
基金supported by the grants from the National Key Research and Development Program of China(2018YFD0201202 and 2017YFD0200601)。
文摘Meloidogyne vitis is a new root-knot nematode parasitic on grape root in Yunnan Province,China.In order to establish a rapid,reliable and specific molecular detection method for M.vitis,the species-specific primers were designed with rDNA-ITS(ribosomal DNA internal transcribed spacer)gene fragment as the target.The reaction system was optimized and the reliability,specificity and sensitivity of primer were testified,therefore,a rapid PCR detection method for M.vitis was established.The result showed that the optimal annealing temperature of the primers was 53℃,which was suitable for the detection of different life stages of M.vitis.Specificity test showed that the specific fragment size of 174 bp was obtained from M.vitis,but other five non-target nematodes did not have any amplification bands,thus effectively distinguish M.vitis and the other five species,and could specifically detect the M.vitis from mixed populations.Sensitivity test showed that this PCR technique could detect the DNA of a single second-stage juvenile(J_(2))and 10^(-4)female.Futhermore,this PCR technique could be used to detect directly M.vitis from soil samples.The rapid,sensitive and specific PCR molecular detection technique could be used for the direct identification of a single J_(2)of M.vitis and the detection of M.vitis in mixed nematode populations and the detection of two J_(2)s or one male in 0.5 g soil samples,which will provide technical support for the investigation of the occurrence and damage of M.vitis and the formulation of efficient green co ntrol strategies.
文摘Enteroaggregative Escherichia coli (EAggEC) strains cause the persistent diarrhea in infants and compromised hosts in developing countries. These strains are currently defined as E. coli that adheres to HEp-2 cells in an aggregative adherence (AA) pattern. In this study, we compared 4 different rapid methods for the detection of EAggEC using a PCR assay, clump formation test, glass slide adherence assay, and the HEp-2 cell adherence assay. Out of 683 E. coli strains isolated from diarrheal stool samples, we detected 17 aggR and/or clump-positive strains, and identified 2 aggR-positive, clump-negative strains and 2 aggR-negative, clump-positive strains. All the aggR positive and clump positive strains also showed positive results in glass slide adherence and HEp-2 cell adherence assays. From all these results, we suggest the following procedure for the rapid identification of EAggEC strains: first, screen E. coli strains with the clump formation test and subsequently perform the glass slide adherence assay to observe AA for confirmation.
文摘Early diagnosis of diseases is critical in its effective management. Traditional disease detection methods require specialized equipment and trained personnel. With the introduction of rapid diagnostic test kits (RDTs), disease detection has become easier and faster. However, these RDTs have failed to compete with the specialized laboratory equipment due to their high detection limits and false alarm rates. This paper presents a novel method of using carbon nanofibers (CNFs) grown on glass microballoons (NMBs) to achieve ultra-low detection limits in RDTs. The NMBs have millions of nanosized CNFs grown on each microballoon, with each CNF having a strong bonding affinity for antibodies. The NMBs conjugated with secondary antibodies have therefore a significantly higher probability of capturing minute antigen concentrations in solution. Furthermore, the dark color formation at the capture zone makes visual disease detection possible. Human Immunoglobulin G (IgG) was selected as the model analyte to study the performance of NMBs using a sandwich immunoassay protocol. Ultra-low electrical detection limit of (4 pg/ml) and rapid re- sponse (~1 minute) was achieved using this method.
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.