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
Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult p...Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult patients with suspected scrub typhus who visited a tertiary care hospital in the Republic of Korea from September to December from 2019 to 2021.The included patients had an acute fever and at least one of the following ten secondary findings:myalgia,skin rash,eschar,headache,thrombocytopenia,increased liver enzyme levels,lymphadenopathy,hepatomegaly,splenomegaly,and pleural effusion.The diagnoses were grouped as scrub typhus or other diseases by two infectious disease physicians.Results:Among 136 patients who met the eligibility criteria,109 had scrub typhus and 27 had different diseases.Single and paired total antibodies using immunofluorescence assay(IFA),and total antibodies using immunochromatography-based rapid diagnostic testing(ICT)were measured in 98%,22%,and 75%of all patients,respectively.Confirmation using paired samples for scrub typhus was established at a median of 11[interquartile range(IQR)10-16]days following the first visit.Among the 82 admitted patients,the median admission time was 9(IQR 7-13)days.According to IFA,58(55%)patients with scrub typhus had total immunoglobulin titers≥1:320,while 23(85%)patients with other disease had titers<1:320.Positive ICT results were observed in 64(74%)patients with scrub typhus and 10(67%)patients with other diseases showed negative ICT results.Conclusions:Serological testing for scrub typhus is currently insufficient for decision-making in clinical practice.展开更多
Background:Swift and accurate detection of Vibrio parahaemolyticus,which is a prominent causative pathogen associated with seafood contamination,is required to effectively combat foodborne disease and wound infections...Background:Swift and accurate detection of Vibrio parahaemolyticus,which is a prominent causative pathogen associated with seafood contamination,is required to effectively combat foodborne disease and wound infections.The toxR gene is relatively conserved within V.parahaemolyticus and is primarily involved in the expression and regulation of virulence genes with a notable degree of specificity.The aim of this study was to develop a rapid,simple,and constant temperature detection method for V.parahaemolyticus in clinical and nonspecialized laboratory settings.Methods:In this study,specific primers and CRISPR RNA were used to target the toxR gene to construct a reaction system that combines recombinase polymerase amplification(RPA)with CRISPR‒Cas13a.The whole-genome DNA of the sample was extracted by self-prepared sodium dodecyl sulphate(SDS)nucleic acid rapid extraction reagent,and visual interpretation of the detection results was performed by lateral flow dipsticks(LFDs).Results:The specificity of the RPA-CRISPR/Cas13a-LFD method was validated using V.parahaemolyticus strain ATCC-17802 and six other non-parahaemolytic Vibrio species.The results demonstrated a specificity of 100%.Additionally,the genomic DNA of V.parahaemolyticus was serially diluted and analysed,with a minimum detectable limit of 1 copy/μL for this method,which was greater than that of the TaqMan-qPCR method(10^(2) copies/μL).The established methods were successfully applied to detect wild-type V.parahaemolyticus,yielding results consistent with those of TaqMan-qPCR and MALDI-TOF MS mass spectrometry identification.Finally,the established RPA-CRISPR/Cas13a-LFD method was applied to whole blood specimens from mice infected with V.parahaemolyticus,and the detection rate of V.parahaemolyticus by this method was consistent with that of the conventional PCR method.Conclusions:In this study,we describe an RPA-CRISPR/Cas13a detection method that specifically targets the toxR gene and offers advantages such as simplicity,rapidity,high specificity,and visual interpretation.This method serves as a valuable tool for the prompt detection of V.parahaemolyticus in nonspecialized laboratory settings.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
[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were ...[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.展开更多
As a marine bacterial pathogen, Photobacterium damselae subsp. damselae(PDD) is distributed in seawater worldwide. It can infect different animals as well as humans, even cause deaths. The highly conserved regions of ...As a marine bacterial pathogen, Photobacterium damselae subsp. damselae(PDD) is distributed in seawater worldwide. It can infect different animals as well as humans, even cause deaths. The highly conserved regions of PDD mcp gene on chromosome and dly gene on plasmid were selected as the target fragments to design the specific primers. Recombinant plasmid standard was prepared based on the primers. With GENECHECKER UF-150 qRT-PCR instrument as the platform, a fluorescence-based quantitative real-time PCR(qRT-PCR) method was established for the detection of PDD. This method can specifically detect PDD and distinguish the highly virulent strains. Additionally, the test results can be qualitatively judged by visualization, while the quantitative detection can be achieved through the standard curve calculation. The minimum limit of detection was 1.0×101 copies μL-1, and the detection time was less than 20 min. In summary, this new method has outstanding advantages, such as strong specificity, high sensitivity, and low site requirements. It is a rapid on-site detection technology for highly virulent PDD strains.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
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
Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to es...Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.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
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
Rice dwarf disease caused by rice dwarf virus （RDV） is one of the major rice virus diseases in China,which widely distributes in the rice area of China.A simple and rapid method for detection of RDV in rice plants a...Rice dwarf disease caused by rice dwarf virus （RDV） is one of the major rice virus diseases in China,which widely distributes in the rice area of China.A simple and rapid method for detection of RDV in rice plants and Nephotettix cincticeps was investigated in this study,and the whole detection only lasted for 20 min.After diseased leaves of rice （10 mg） and leaves of Nephotettix cincticeps （10 mg） both infected by RDV were ground by sterile water （100 μl）,the supernatant was analyzed by 1% agarose gel electrophoresis to steadily detect five specific electrophoretic bands with 1 000 bp to 5 000 bp.However,there was no electrophoretic band in the healthy rice leaves and aqueous extract of non-viruliferous N.cincticeps.Therefore,this method was more rapid and simple to detect RDV in rice plants and Nephotettix cincticeps,which avoided expensive reagents and tedious steps of conventional serological testing and molecular detection （RT-PCR and Western-blot）.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
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
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
Numerous large-scale fragmented bedrock landslides developed along major fault system is a world-wide phenomenon,which are often characterized with repeated reactivation throughout histories.Due to the large-scale and...Numerous large-scale fragmented bedrock landslides developed along major fault system is a world-wide phenomenon,which are often characterized with repeated reactivation throughout histories.Due to the large-scale and deep-seated features,it is normally difficult to control such landslides,which in turn pose great threat to local residents and infrastructures.Therefore,monitoring and forecasting these gigantic landslides has become a key protocol for risk reduction.This paper introduces such a typical massive landslide,named Yahuokou landslide,besides Min River in Zhouqu County,Gansu Province,China.Reactivated on July 16,2019 with a volume of approximately 4×106 m3,moving slowly and transitionally starting from top part,its toe had partially blocked the Min River and destroyed roads and houses eventually by August 11,2019.As to emergency response for such huge slowmoving landslide,there is no standard national protocols.Therefore,how to make effective emergency decision has become a challenge.Based on previous experiences,integrated multi-methods,including UAV imagery interpretation,we applied GNSS monitoring and field investigations in the early stages of landsliding,in order to assist the decisionmaking.The results show that the movement path of the current displacement is consistent with that of the 1989 reactivation event,and the slide body was separated into three relatively independent blocks with different sliding velocities and responses to rainfall.The upper and lower blocks appeared less affected by rainfall,while the middle block responded more to the changes in precipitations.It proves that the combined approaches using a variety of monitoring techniques can play an effective role in the monitoring of rapidly deformed transitional largescale landslides,and can also provide a set of reference methods for the emergency disposal of similar landslide hazards.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
Mycotoxins are secondary metabolites produced by fungus.Many mycotoxin species are highly toxic and are frequently found in cereals and feedstuffs.So,powerful detection methods are vital and effective ways to prevent ...Mycotoxins are secondary metabolites produced by fungus.Many mycotoxin species are highly toxic and are frequently found in cereals and feedstuffs.So,powerful detection methods are vital and effective ways to prevent feed contamination.Traditional detection methods can no longer meet the needs of massive,real-time,simple,and fast mycotoxin monitoring.Rapid detection methods based on advanced material and sensor technology are the future trend.In this review,we highlight recent progress of mycotoxin rapid detection strategies in feedstuffs and foods,especially for simultaneous multiplex mycotoxin determination.Immunoassays,biosensors,and the prominent roles of nanomaterials are introduced.The principles of different types of recognition and signal transduction are explained,and the merits and pitfalls of these methods are compared.Furthermore,limitations and challenges of existing rapid sensing strategies and perspectives of future research are discussed.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
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the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI),funded by the Ministry of Health&Welfare,Republic of Korea(grant no.HI22C0306).
文摘Objective:Serological tests are widely used for scrub typhus diagnosis;however,their limitations are evident.This study aims to assess their practical value in clinical settings.Methods:We analyzed the data of adult patients with suspected scrub typhus who visited a tertiary care hospital in the Republic of Korea from September to December from 2019 to 2021.The included patients had an acute fever and at least one of the following ten secondary findings:myalgia,skin rash,eschar,headache,thrombocytopenia,increased liver enzyme levels,lymphadenopathy,hepatomegaly,splenomegaly,and pleural effusion.The diagnoses were grouped as scrub typhus or other diseases by two infectious disease physicians.Results:Among 136 patients who met the eligibility criteria,109 had scrub typhus and 27 had different diseases.Single and paired total antibodies using immunofluorescence assay(IFA),and total antibodies using immunochromatography-based rapid diagnostic testing(ICT)were measured in 98%,22%,and 75%of all patients,respectively.Confirmation using paired samples for scrub typhus was established at a median of 11[interquartile range(IQR)10-16]days following the first visit.Among the 82 admitted patients,the median admission time was 9(IQR 7-13)days.According to IFA,58(55%)patients with scrub typhus had total immunoglobulin titers≥1:320,while 23(85%)patients with other disease had titers<1:320.Positive ICT results were observed in 64(74%)patients with scrub typhus and 10(67%)patients with other diseases showed negative ICT results.Conclusions:Serological testing for scrub typhus is currently insufficient for decision-making in clinical practice.
文摘Background:Swift and accurate detection of Vibrio parahaemolyticus,which is a prominent causative pathogen associated with seafood contamination,is required to effectively combat foodborne disease and wound infections.The toxR gene is relatively conserved within V.parahaemolyticus and is primarily involved in the expression and regulation of virulence genes with a notable degree of specificity.The aim of this study was to develop a rapid,simple,and constant temperature detection method for V.parahaemolyticus in clinical and nonspecialized laboratory settings.Methods:In this study,specific primers and CRISPR RNA were used to target the toxR gene to construct a reaction system that combines recombinase polymerase amplification(RPA)with CRISPR‒Cas13a.The whole-genome DNA of the sample was extracted by self-prepared sodium dodecyl sulphate(SDS)nucleic acid rapid extraction reagent,and visual interpretation of the detection results was performed by lateral flow dipsticks(LFDs).Results:The specificity of the RPA-CRISPR/Cas13a-LFD method was validated using V.parahaemolyticus strain ATCC-17802 and six other non-parahaemolytic Vibrio species.The results demonstrated a specificity of 100%.Additionally,the genomic DNA of V.parahaemolyticus was serially diluted and analysed,with a minimum detectable limit of 1 copy/μL for this method,which was greater than that of the TaqMan-qPCR method(10^(2) copies/μL).The established methods were successfully applied to detect wild-type V.parahaemolyticus,yielding results consistent with those of TaqMan-qPCR and MALDI-TOF MS mass spectrometry identification.Finally,the established RPA-CRISPR/Cas13a-LFD method was applied to whole blood specimens from mice infected with V.parahaemolyticus,and the detection rate of V.parahaemolyticus by this method was consistent with that of the conventional PCR method.Conclusions:In this study,we describe an RPA-CRISPR/Cas13a detection method that specifically targets the toxR gene and offers advantages such as simplicity,rapidity,high specificity,and visual interpretation.This method serves as a valuable tool for the prompt detection of V.parahaemolyticus in nonspecialized laboratory settings.
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.
基金Supported by The National Project for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39).
文摘[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.
基金supported by the National Key Research and Development Program of China (No. 2019YFD0900104)the Key Projects of Science and Technology In-novation of Shandong Province (No. 2018YFJH0703)。
文摘As a marine bacterial pathogen, Photobacterium damselae subsp. damselae(PDD) is distributed in seawater worldwide. It can infect different animals as well as humans, even cause deaths. The highly conserved regions of PDD mcp gene on chromosome and dly gene on plasmid were selected as the target fragments to design the specific primers. Recombinant plasmid standard was prepared based on the primers. With GENECHECKER UF-150 qRT-PCR instrument as the platform, a fluorescence-based quantitative real-time PCR(qRT-PCR) method was established for the detection of PDD. This method can specifically detect PDD and distinguish the highly virulent strains. Additionally, the test results can be qualitatively judged by visualization, while the quantitative detection can be achieved through the standard curve calculation. The minimum limit of detection was 1.0×101 copies μL-1, and the detection time was less than 20 min. In summary, this new method has outstanding advantages, such as strong specificity, high sensitivity, and low site requirements. It is a rapid on-site detection technology for highly virulent PDD strains.
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.
基金Supported by the Natural Science Foundation of Heilongjiang Province(Topic C2017032)Heilongjiang Province Applied Technology Research and Development Program(Topic GA19B104)the National Key Research and Development Program(Topic 2018YFD0300105)。
文摘Rice bacterial leaf brown spot disease caused by Pseudomonas syringae pv.syringae(Pss)is a major disease on rice.In recent years,Pss has emerged worldwide,seriously affecting rice production.It is very important to establish a rapid detection method of Pss for the diagnosis and prevention of this disease.In order to robust and accurately diagnose the rice bacterial leaf brown spot disease in the field and laboratory,an assay system for the Pss was developed in this study,and the specific sequence of hrcN was used as the target,based on loop-mediated isothermal amplification(LAMP).The best detection system was MgSO 48 mmol·L^(-1),Bst DNA polymerase 8 U,dNTP 1.4 mmol·L^(-1),the ratio of internal and outer primers was 2:1,the reaction temperature was 63℃,the reaction time was 45 min,and the lowest sensitivity was 104 CFU·mL^(-1).This results provided an accurate and robust method for laboratory and field diagnosis of bacterial leaf brown spot disease of rice.
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.
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Rice dwarf disease caused by rice dwarf virus （RDV） is one of the major rice virus diseases in China,which widely distributes in the rice area of China.A simple and rapid method for detection of RDV in rice plants and Nephotettix cincticeps was investigated in this study,and the whole detection only lasted for 20 min.After diseased leaves of rice （10 mg） and leaves of Nephotettix cincticeps （10 mg） both infected by RDV were ground by sterile water （100 μl）,the supernatant was analyzed by 1% agarose gel electrophoresis to steadily detect five specific electrophoretic bands with 1 000 bp to 5 000 bp.However,there was no electrophoretic band in the healthy rice leaves and aqueous extract of non-viruliferous N.cincticeps.Therefore,this method was more rapid and simple to detect RDV in rice plants and Nephotettix cincticeps,which avoided expensive reagents and tedious steps of conventional serological testing and molecular detection （RT-PCR and Western-blot）.
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 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.
基金funded by National Key Research and Development Program of China(Grant No.2017YFC1501005)the National Key R&D Program of China(Grant No.2018YFC1504704)+3 种基金the National Natural Science Foundation of China(Grant No.42007232)the Key Research and Development Program of Gansu Province(Grant No.20YF8FA074)the Science and Technology Major Project of Gansu Province(Grant No.19ZD2FA002)the Construction Project of Gansu Technological Innovation Center(Grant No.18JR2JA006)。
文摘Numerous large-scale fragmented bedrock landslides developed along major fault system is a world-wide phenomenon,which are often characterized with repeated reactivation throughout histories.Due to the large-scale and deep-seated features,it is normally difficult to control such landslides,which in turn pose great threat to local residents and infrastructures.Therefore,monitoring and forecasting these gigantic landslides has become a key protocol for risk reduction.This paper introduces such a typical massive landslide,named Yahuokou landslide,besides Min River in Zhouqu County,Gansu Province,China.Reactivated on July 16,2019 with a volume of approximately 4×106 m3,moving slowly and transitionally starting from top part,its toe had partially blocked the Min River and destroyed roads and houses eventually by August 11,2019.As to emergency response for such huge slowmoving landslide,there is no standard national protocols.Therefore,how to make effective emergency decision has become a challenge.Based on previous experiences,integrated multi-methods,including UAV imagery interpretation,we applied GNSS monitoring and field investigations in the early stages of landsliding,in order to assist the decisionmaking.The results show that the movement path of the current displacement is consistent with that of the 1989 reactivation event,and the slide body was separated into three relatively independent blocks with different sliding velocities and responses to rainfall.The upper and lower blocks appeared less affected by rainfall,while the middle block responded more to the changes in precipitations.It proves that the combined approaches using a variety of monitoring techniques can play an effective role in the monitoring of rapidly deformed transitional largescale landslides,and can also provide a set of reference methods for the emergency disposal of similar landslide hazards.
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 financial support from the National Key Research and Development Program of China(2017YFC1600300).
文摘Mycotoxins are secondary metabolites produced by fungus.Many mycotoxin species are highly toxic and are frequently found in cereals and feedstuffs.So,powerful detection methods are vital and effective ways to prevent feed contamination.Traditional detection methods can no longer meet the needs of massive,real-time,simple,and fast mycotoxin monitoring.Rapid detection methods based on advanced material and sensor technology are the future trend.In this review,we highlight recent progress of mycotoxin rapid detection strategies in feedstuffs and foods,especially for simultaneous multiplex mycotoxin determination.Immunoassays,biosensors,and the prominent roles of nanomaterials are introduced.The principles of different types of recognition and signal transduction are explained,and the merits and pitfalls of these methods are compared.Furthermore,limitations and challenges of existing rapid sensing strategies and perspectives of future research are discussed.
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.
