Venereal diseases are considered to be the most prevalent infectious diseases in the worldwide. China is now faced with a year-by-year increasing incidence of sexually transmitted diseases （STD）, which are spreading...Venereal diseases are considered to be the most prevalent infectious diseases in the worldwide. China is now faced with a year-by-year increasing incidence of sexually transmitted diseases （STD）, which are spreading from high-risk groups to the general population. Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum and herpes simplex virus-2 （HSV-2） are always regarded as the most common venereal pathogens. The ＂golden standard＂ for testing Neisseria gonorrhoeae remains to be bacteria culture or microscopic examination.展开更多
Objective: to analyze the advantages of microbial testing. Methods: a total of 200 patients with urinary system infection were selected and divided into two groups, and different testing methods were used. Results: th...Objective: to analyze the advantages of microbial testing. Methods: a total of 200 patients with urinary system infection were selected and divided into two groups, and different testing methods were used. Results: the detection rate of pathogenic bacteria in the observation group was 90.00% higher than that in the control group, which was 60.00% (P<0.05). Conclusion: microbial testing has many advantages, which can not only ensure the curative effect of patients, but also prevent and reduce the occurrence of infection risk in patients.展开更多
While the pathogen nucleic acid diagnostic technology has made tremendous progresses,several challenges,including long development cycles and limited specificity still exist,especially in the context of isothermal amp...While the pathogen nucleic acid diagnostic technology has made tremendous progresses,several challenges,including long development cycles and limited specificity still exist,especially in the context of isothermal amplification techniques.To enhance the detection accuracy,here a functional strand displacement catalytic hairpin assembly circuit,which can perform at high-temperature(HT-CHA),was developed as the downstream of the loop mediated isothermal nucleic acid amplification(LAMP).The addition of HT-CHA not only ensures the specificity but also amplifies the detection signal.Taking African swine fever(ASF)gene as the target,the input of HT-CHA was designed according to the ASF gene LAMP amplicons.This LAMP-HTCHA can detect as low as 2 copies/μL targeting genes with high signal-to-noise ratio.Through importing a three-way junction(3WJ)transducer,the HT-CHA well-developed for ASF gene can be directly adapted to detect the LAMP amplicons of other pathogen genes,such as Mycoplasma pneumoniae(MP)gene.The time-consuming and high-risk process to redesign HT-CHA components can be further avoided,making the method even general and useful for a plenty of other targets.Finally,the accurate detection of MP gene in alveolar lavage fluid samples confirmed the high potential of the LAMP and HT-CHA combination in clinical applications,providing a promising strategy to develop point-of-care diagnostics at constant temperatures.展开更多
The COVID-19 pandemic evidenced the urgent need for rapid,accurate,and scalable diagnostic methods for emerging infectious diseases.Droplet digital reverse transcription LAMP(ddRT-LAMP)is a promising technique for pat...The COVID-19 pandemic evidenced the urgent need for rapid,accurate,and scalable diagnostic methods for emerging infectious diseases.Droplet digital reverse transcription LAMP(ddRT-LAMP)is a promising technique for pathogen detection and accurate quantification,as it overcomes traditional LAMP’s limitations in viral load estimation through reaction partitioning and digital analysis.However,many parameters must be adjusted to avoid spurious results.This study evaluates the critical conditions for effective ddRT-LAMP quantification of the SARS-CoV-2 N gene in plasmid DNA,synthetic RNA,and nasopharyngeal swab samples.Using a polydimethylsiloxane(PDMS)microfluidic device,the RT-LAMP reaction mixture with a fluorescent dye was divided into thousands of droplets stabilized by a surfactant in fluorinated oil.After incubation,the droplets were injected into a PDMS chamber for fluorescent imaging to determine the proportion of positive droplets and quantify the samples based on the Poisson distribution.The results showed that primer design and master mix composition significantly impacted the amplification.The selection of GelGreen^(R)as the fluorescent dye was crucial,as other dyes tested diffused into the oil phase.Optimal amplification occurred with 105μm droplet diameter and 30-min incubation,achieving detection and quantification limits of 102 cp/μL.By addressing these operational challenges,ddRT-LAMP can become a more effective tool for viral detection and quantification in clinical diagnostics.展开更多
Pathogens pose significant threats to biosecurity and environmental health due to their potential for widespread outbreaks.Effective pathogen detection requires methods that are rapid,sensitive,specific,and informativ...Pathogens pose significant threats to biosecurity and environmental health due to their potential for widespread outbreaks.Effective pathogen detection requires methods that are rapid,sensitive,specific,and informative.Here,we proposed a multiplex visual detection system that integrated ultrafast polymerase chain reaction(PCR)and molecular beacons,allowing the simultaneous detection of three pathogens in a one-pot reaction.The ultrafast PCR protocol employed cycles of just 7 s each,allowing the entire process-from sampling to result-to be completed within only 10 min.Molecular beacons hybridized with target sequences during ultrafast PCR,generating fluorescence signals that are visually detectable without specialized equipment.Additionally,we developed a compact,portable cartridge integrated with online software for fluorescence visualization and direct result output,eliminating the need for bulky instruments and specialized personnel,thereby facilitating point-of-care testing(POCT).The method demonstrated high specificity and sensitivity,with a limit of detection(LOD)as low as 23 copies per reaction.It achieved a 100%positive detection rate in practical applications,performing comparably to standard PCR.Furthermore,the method effectively identified low concentrations of pathogens in animal infection samples.This ultrafast,highly sensitive,specific,and informative method shows significant potential for POCT applications,including food safety monitoring and clinical diagnostics.展开更多
Conjugated polymers （CPs） are referred to a kind of fluorescent polymer materials with delocalized n-conjugated backbones. For the last decades, cationic CPs （CCPs） have been widely used in biosensor, imaging and ...Conjugated polymers （CPs） are referred to a kind of fluorescent polymer materials with delocalized n-conjugated backbones. For the last decades, cationic CPs （CCPs） have been widely used in biosensor, imaging and biomedical fields due to their good photophysical properties and solubility in water medium resulting from side chain modification with ionized moieties. In this mini-review, we mainly introduced the applications of CCPs in detection and inactivation of pathogen with typical examples, and also briefly discussed the relevant mechanisms. We hold the expectation that this mini-review can offer researchers a general reference and inspire them to construct new systems with high performances of pathogen detection and antimicrobial activity.展开更多
Infectious disease outbreaks have seriously endangered global health owing to the scarcity of testing materials and techniques.Diversified materials and methods should be urgently developed for rapid detection and dis...Infectious disease outbreaks have seriously endangered global health owing to the scarcity of testing materials and techniques.Diversified materials and methods should be urgently developed for rapid detection and discrimination of pathogenic microorganisms.Conjugated polymer(CP)materials are macromolecular compounds comprising numerous covalently bonded luminescent units.They have excellent light-harvesting and optical signal amplification capabilities owing to the transmission of excitation energy along their backbone.In recent years,CP materials have aroused research enthusiasm in the biosensors field because of their outstanding optoelectronic properties.This brief manuscript provides an overall review of recent progress achieved in CP-based systems for pathogen sensing.展开更多
Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg...Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg-F1;R2)was designed based on the RPB sequence,and real-time fluorescence quantitative PCR(qPCR)was used to validate the efficiency of the primer.The results showed that the primer pair had high specificity and sensitivity of 100 pg of DNA.Furthermore,the qPCR system for early and rapid detection of F.peasudeograminearum had an amplification efficiency of 87.5%and correlation coefficient of 0.99,and the pathologic threshold of F.pseudograminearum in soil was determined by using this detection system.It was found that F.pseudograminearum could cause Fusarium crown rot when the DNA concentration of F.pseudograminearum in field soil exceeded 213 pg·g^(-1).Hence,the qPCR-based method we developed for F.pseudograminearum detection has the advantages of high specificity and sensitivity,and can be used for rapid and early detection of F.pseudograminearum even in field soils.展开更多
In recent years,with the rapid development of molecular biology diagnostic technology,many new polymerase chain reaction(PCR)technologies with high specificity and good sensitivity have gradually been developed.While ...In recent years,with the rapid development of molecular biology diagnostic technology,many new polymerase chain reaction(PCR)technologies with high specificity and good sensitivity have gradually been developed.While expanding the range of detection methods,these technologies inevitably have some disadvantages.Therefore,in clinical pathogen diagnosis,medical personnel should choose the detection method according to the detection purpose and pathogen characteristics.In this paper,the basic principle,application scope,advantages and disadvantages and development of various emerging PCR diagnostic techniques are respectively described in order to provide a theoretical reference for the selection of pathogenic biological diagnostic techniques in the clinical practice.展开更多
To the Editor:Multi-pathogen detection has become increasingly important,given the increasing risk of circulating contagious pathogens due to global changes characterized by urbanization and habitat fragmentation.The ...To the Editor:Multi-pathogen detection has become increasingly important,given the increasing risk of circulating contagious pathogens due to global changes characterized by urbanization and habitat fragmentation.The coronavirus disease-2019(COVID-19)pandemic disrupted the epidemiology of some respiratory pathogens,possibly due to public health and social measures,as well as changes in healthcare-seeking behaviors and interactions between viruses.Multi-respiratory pathogen surveillance could be useful for adjusting disease prevention strategies and preventing potential outbreaks of existing and emerging respiratory infectious diseases.On the other hand,symptoms and disease severity differ among respiratory pathogen infections,[1]and coinfection might be associated with increased disease severity.[2]Some respiratory pathogens such as severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),influenza viruses(IFV),respiratory syncytial virus(RSV),and Streptococcus pneumoniae(S.pneumoniae)impose a high disease burden on populations but can be prevented by vaccines.Therefore,this study aimed to determine clinical manifestations and disease severity among patients testing positive for various pathogens to offer guidance on disease diagnosis,treatment,surveillance,and immunization strategies.展开更多
Rare infectious diseases are infections that are uncommon,have a low incidence,and are caused by newly emerging pathogens,cross‐species or ectopic infections,or host immunodeficiencies.The detection and diagnosis of ...Rare infectious diseases are infections that are uncommon,have a low incidence,and are caused by newly emerging pathogens,cross‐species or ectopic infections,or host immunodeficiencies.The detection and diagnosis of rare infections is one of the main reasons for misdiagnosis and missed diagnosis.Traditional detection techniques such as microbial cultivation and isolation or polymerase chain reaction often fail to meet the clinical demands for timeliness,accuracy,and sensitivity.Metagenomic nextgeneration sequencing involves pan-nucleic acid testing conducted directly using specimens to facilitate rapid identification of rare or unidentified pathogens.Despite the availability of various techniques,advanced methods in clinical practice are necessary to achieve timely clinical diagnosis of rare infections.In this review,we summarize the definition and clinical significance of rare infectious diseases as well as the current detection methods,limitations,and future research areas for their detection.展开更多
[Objectives]This study was conducted to evaluate the immunization efficacy and infection status of classical swine fever(CSF),foot-and-mouth disease(FMD),porcine reproductive and respiratory syndrome(PRRS),pseudorabie...[Objectives]This study was conducted to evaluate the immunization efficacy and infection status of classical swine fever(CSF),foot-and-mouth disease(FMD),porcine reproductive and respiratory syndrome(PRRS),pseudorabies(PR),and porcine circovirus type 2(PCV2)in large-scale pig farms.[Methods]Antibody and pathogen detection was performed on 56 serum samples collected in March 2025.[Results]The antibody qualification rates for CSF,FMD,and PRRS were 76.8%,73.2%,and 76.8%,respectively,all meeting the national standards.However,nursery pigs exhibited an immunity gap,indicating a need for timely booster vaccinations.No PRV gE antibodies or PCV2 antibodies were detected,reflecting the absence of vaccination against these diseases and suggesting significant effectiveness of comprehensive biosecurity measures.The low antibody qualification rate for PRRS in the nursery stage highlights the need for improved immunization management.[Conclusions]This study provides data support and practical insights for integrated disease prevention and control in large-scale pig farms.展开更多
The most widely used method of identification of microbial morphology and structure is microscopy,but it can be difficult to distinguish between pathogens with a similar appearance.Existing fluorescent staining method...The most widely used method of identification of microbial morphology and structure is microscopy,but it can be difficult to distinguish between pathogens with a similar appearance.Existing fluorescent staining methods require a combination of a variety of fluorescent materials to meet this demand.In this study,unique concentration-dependent fluorescent carbon dots(CDs)were synthesized for the identification and quantification of pathogens.The emission wavelength of the CDs could be tuned spanning the full visible region by virtue of aggregation-induced narrowing of bandgaps.This tunable emission wavelength of the specific concentration response to diverse microbes can be used to distinguish microorganisms with a similar appearance,even in a same genus.A hyperspectral microscopy system was demonstrated to distinguish Aspergillus flavus and A.fumigatus based on the results above.The identification accuracy of the two similar-looking pathogens can be close to 100%,and the relative proportions and spatial distributions can also be profiled from the mixture of the pathogens.This technique can provide a solution to the fast detection of microorganisms and is potentially applicable to a wide range of problems in areas such as healthcare,food preparation,biotechnology,and health emergency.展开更多
Rapid,high-throughput,timely,multiplex diagnosis of respiratory-tract infections still relies on laboratory infrastructure,sequential assays,and trained personnel,thereby delaying targeted therapy and outbreak contain...Rapid,high-throughput,timely,multiplex diagnosis of respiratory-tract infections still relies on laboratory infrastructure,sequential assays,and trained personnel,thereby delaying targeted therapy and outbreak containment.In this study,a Fully Automated rotary microfluidic platform(FA-RMP)for high-throughput multiplex respiratory tract pathogens detection was presented.FA-RMP enables a true“sample-in,result-out”workflow through the integration of swab lysis,reagent partitioning,lyophilized reverse transcription loop-mediated isothermal amplification(RT-LAMP),and movingprobe fluorescence read-out,all encapsulated with a disposable microfluidic cartridge and paired with a 9 kg,fourchannel benchtop reader.The FA-RMP enables parallel processing of 16 independent reactions within 30 min,supporting simultaneous detection of up to 4 distinct clinical samples.Analytical validation using serially diluted Mycoplasma pneumoniae(MP)DNA established a limit of detection(LoD)of 50 copies μL^(-1) and a log-linear correlation between threshold time and template load(R^(2)=0.9528).Testing with eight non-target respiratory pathogens yielded no amplification,confirming high analytical specificity.FA-RMP successfully detected the clinical samples with influenza A,influenza B,and MP,further demonstrating its robust multiplex detection capability.By integrating automated sample preparation,multiplex isothermal amplification and quantitative detection into a portable,high-throughput system,the platform delivers laboratory-grade performance at the point of care,serving as a scalable tool for routine respiratory pathogens screening and rapid epidemic response.展开更多
Introduction:Fluorescent probe-based recombinase aided amplification(RAA)offers the advantages of rapidity and simplicity but is limited by the requirement for complex and lengthy probe design,restricting its widespre...Introduction:Fluorescent probe-based recombinase aided amplification(RAA)offers the advantages of rapidity and simplicity but is limited by the requirement for complex and lengthy probe design,restricting its widespread application.Methods:A novel EvaGreen dye-based RAA(EvaGreen-RAA)assay utilizing self-avoiding molecular recognition system(SAMRS)primers was developed for the detection of Pseudomonas fluorescens(PF)and Bacillus cereus(BC)in milk.Conventional RAA was used as a reference method.Sensitivity was evaluated using nucleic acids from recombinant plasmids and simulated milk specimens.Additionally,a dual EvaGreen-RAA assay was investigated for simultaneous detection of mixed BC and PF in simulated milk specimens.Results:The EvaGreen-RAA demonstrated superior sensitivity compared to conventional RAA,with detection limits of 1 copy/μL versus 10 copies/μL for both BC and PF plasmids,respectively.In simulated milk specimens,EvaGreen-RAA detected BC and PF at concentrations of 100 CFU/mL and 200 CFU/mL,respectively,compared to 400 CFU/mL and 600 CFU/mL for conventional RAA.The dual EvaGreen-RAA assay successfully detected mixed BC and PF in simulated milk specimens at concentrations of 200 CFU/mL for each pathogen.Conclusion:The EvaGreen-RAA assay demonstrated significant advantages in terms of simplicity and enhanced sensitivity compared to fluorescent probe-based RAA,offering a novel approach for developing multiplex pathogen detection systems using melting curve analysis.展开更多
In the food production sector,quickly identifying potential hazards is crucial due to the resilience of many pathogens,which could lead to wasted production results and,more severely,epidemic outbreaks.E.coli monitori...In the food production sector,quickly identifying potential hazards is crucial due to the resilience of many pathogens,which could lead to wasted production results and,more severely,epidemic outbreaks.E.coli monitoring is essential;however,traditional quality control methods in fish farming are often slow and intrusive,thus promoting an increase in fish stress and mortality rates.This paper presents an alternative method by utilizing a prototype inspired by polarized optical microscopy(POM),constructed with a Raspberry Pi microprocessor to assess pixel patterns and calculate analyte levels.展开更多
Some plants use intracellular immune receptors,mainly from the nucleotide-binding leucine-rich repeat receptors(NLRs)super-family,to detect pathogen effectors and restrict pathogen infection.NLRs are categorized as TI...Some plants use intracellular immune receptors,mainly from the nucleotide-binding leucine-rich repeat receptors(NLRs)super-family,to detect pathogen effectors and restrict pathogen infection.NLRs are categorized as TIR-NLR(TNL)or CC-NLR(CNL)based on their N-terminal Tollinterleukin 1-like receptor(TIR)or coiled-coil(CC)domain.Additionally,there are helper(h)NLRs that aid in translating signals from pathogen-sensing NLRs into effector-triggered immunity responses.NLRs can directly detect effectors or indirectly monitor their host targets,leading to a robust immune response and programmed cell death,known as the hypersensitive response(HR).展开更多
A sustainable early warning monitoring system can overcome key limitations of conventional approaches for managing infectious diseases and epidemics.It enables consistent monitoring of pathogens,including SARSCoV-2,in...A sustainable early warning monitoring system can overcome key limitations of conventional approaches for managing infectious diseases and epidemics.It enables consistent monitoring of pathogens,including SARSCoV-2,in wastewater treatment plants(WWTPs).Wastewater-based epidemiology(WBE)analyses biological markers excreted in sewage to monitor community health.It has emerged as a powerful tool because it is cost-effective,non-invasive,and capable of detecting pathogen circulation within the population,even before clinical cases are reported[1].These features provide WBE a clear advantage over traditional clinical testing,particularly by enabling the early detection of new variants[2].展开更多
Sensory mechanisms play pivotal roles in plant defense against pathogen infection.To safeguard themselves,plants developed both cell surface-situated pattern recognition receptors(PRRs)to identify pathogen-associated ...Sensory mechanisms play pivotal roles in plant defense against pathogen infection.To safeguard themselves,plants developed both cell surface-situated pattern recognition receptors(PRRs)to identify pathogen-associated molecular patterns(PAMPs)and intracellular nucleotidebinding leucine-rich repeat receptors(NLRs)to detect pathogen effectors when they face imminent threats from a wide range of pathogens(Boller and Felix,22009:Jones et al.,2024;Wu et al.,2014).展开更多
基金supported by Zhejiang Provincial Population and Family Planning Foundation of China (N20100011)
文摘Venereal diseases are considered to be the most prevalent infectious diseases in the worldwide. China is now faced with a year-by-year increasing incidence of sexually transmitted diseases （STD）, which are spreading from high-risk groups to the general population. Neisseria gonorrhoeae, Chlamydia trachomatis, Ureaplasma urealyticum and herpes simplex virus-2 （HSV-2） are always regarded as the most common venereal pathogens. The ＂golden standard＂ for testing Neisseria gonorrhoeae remains to be bacteria culture or microscopic examination.
文摘Objective: to analyze the advantages of microbial testing. Methods: a total of 200 patients with urinary system infection were selected and divided into two groups, and different testing methods were used. Results: the detection rate of pathogenic bacteria in the observation group was 90.00% higher than that in the control group, which was 60.00% (P<0.05). Conclusion: microbial testing has many advantages, which can not only ensure the curative effect of patients, but also prevent and reduce the occurrence of infection risk in patients.
基金the support of the Natural Science Foundation of China(22004118)Key R&D Program of Jilin Province(20230203193SF)Cooperation funding of Changchun with Chinese academy of sciences(21SH16).
文摘While the pathogen nucleic acid diagnostic technology has made tremendous progresses,several challenges,including long development cycles and limited specificity still exist,especially in the context of isothermal amplification techniques.To enhance the detection accuracy,here a functional strand displacement catalytic hairpin assembly circuit,which can perform at high-temperature(HT-CHA),was developed as the downstream of the loop mediated isothermal nucleic acid amplification(LAMP).The addition of HT-CHA not only ensures the specificity but also amplifies the detection signal.Taking African swine fever(ASF)gene as the target,the input of HT-CHA was designed according to the ASF gene LAMP amplicons.This LAMP-HTCHA can detect as low as 2 copies/μL targeting genes with high signal-to-noise ratio.Through importing a three-way junction(3WJ)transducer,the HT-CHA well-developed for ASF gene can be directly adapted to detect the LAMP amplicons of other pathogen genes,such as Mycoplasma pneumoniae(MP)gene.The time-consuming and high-risk process to redesign HT-CHA components can be further avoided,making the method even general and useful for a plenty of other targets.Finally,the accurate detection of MP gene in alveolar lavage fluid samples confirmed the high potential of the LAMP and HT-CHA combination in clinical applications,providing a promising strategy to develop point-of-care diagnostics at constant temperatures.
基金preliminary LAMP experiments in bulk by Dr.Elizabeth Castillo-Villanueva.K.C.R.acknowledges Secretaría de Ciencia,Humanidades,Tecnología e Innovación(SECIHTI),for a postdoctoral scholarship(CVU 662463)supported by UNAM-PAPIIT grants IT100922 and IN211623,SECTEI grant SECTEI/248/2021the Faculty of Chemistry,UNAM(PAIP 5000-9023).
文摘The COVID-19 pandemic evidenced the urgent need for rapid,accurate,and scalable diagnostic methods for emerging infectious diseases.Droplet digital reverse transcription LAMP(ddRT-LAMP)is a promising technique for pathogen detection and accurate quantification,as it overcomes traditional LAMP’s limitations in viral load estimation through reaction partitioning and digital analysis.However,many parameters must be adjusted to avoid spurious results.This study evaluates the critical conditions for effective ddRT-LAMP quantification of the SARS-CoV-2 N gene in plasmid DNA,synthetic RNA,and nasopharyngeal swab samples.Using a polydimethylsiloxane(PDMS)microfluidic device,the RT-LAMP reaction mixture with a fluorescent dye was divided into thousands of droplets stabilized by a surfactant in fluorinated oil.After incubation,the droplets were injected into a PDMS chamber for fluorescent imaging to determine the proportion of positive droplets and quantify the samples based on the Poisson distribution.The results showed that primer design and master mix composition significantly impacted the amplification.The selection of GelGreen^(R)as the fluorescent dye was crucial,as other dyes tested diffused into the oil phase.Optimal amplification occurred with 105μm droplet diameter and 30-min incubation,achieving detection and quantification limits of 102 cp/μL.By addressing these operational challenges,ddRT-LAMP can become a more effective tool for viral detection and quantification in clinical diagnostics.
基金the National Natural Science Foundation of China(32371521,32172285,82300220)Special Project for Experimental Animal Research(23141900300)+4 种基金the Foundation for Scholars of Fuzhou University(XRC-24068)Shanghai Rising Star Program(23QA1404300)Special Project for Medical Innovation Research(22Y11909200)Greater Bay Area Institute of Precision Medicine(Guangzhou)Human Phenome Data Center of Fudan University and Shanghai Municipal Science and Technology Major Project(2023SHZDZX02).
文摘Pathogens pose significant threats to biosecurity and environmental health due to their potential for widespread outbreaks.Effective pathogen detection requires methods that are rapid,sensitive,specific,and informative.Here,we proposed a multiplex visual detection system that integrated ultrafast polymerase chain reaction(PCR)and molecular beacons,allowing the simultaneous detection of three pathogens in a one-pot reaction.The ultrafast PCR protocol employed cycles of just 7 s each,allowing the entire process-from sampling to result-to be completed within only 10 min.Molecular beacons hybridized with target sequences during ultrafast PCR,generating fluorescence signals that are visually detectable without specialized equipment.Additionally,we developed a compact,portable cartridge integrated with online software for fluorescence visualization and direct result output,eliminating the need for bulky instruments and specialized personnel,thereby facilitating point-of-care testing(POCT).The method demonstrated high specificity and sensitivity,with a limit of detection(LOD)as low as 23 copies per reaction.It achieved a 100%positive detection rate in practical applications,performing comparably to standard PCR.Furthermore,the method effectively identified low concentrations of pathogens in animal infection samples.This ultrafast,highly sensitive,specific,and informative method shows significant potential for POCT applications,including food safety monitoring and clinical diagnostics.
基金supported by the National Natural Science Foundation of China (21473221, 21473220)
文摘Conjugated polymers （CPs） are referred to a kind of fluorescent polymer materials with delocalized n-conjugated backbones. For the last decades, cationic CPs （CCPs） have been widely used in biosensor, imaging and biomedical fields due to their good photophysical properties and solubility in water medium resulting from side chain modification with ionized moieties. In this mini-review, we mainly introduced the applications of CCPs in detection and inactivation of pathogen with typical examples, and also briefly discussed the relevant mechanisms. We hold the expectation that this mini-review can offer researchers a general reference and inspire them to construct new systems with high performances of pathogen detection and antimicrobial activity.
基金supported by the Natural Science Foundation of China(No.21704005)Fundamental Research Funds for the Cen-tral Universities(No.300102310106).
文摘Infectious disease outbreaks have seriously endangered global health owing to the scarcity of testing materials and techniques.Diversified materials and methods should be urgently developed for rapid detection and discrimination of pathogenic microorganisms.Conjugated polymer(CP)materials are macromolecular compounds comprising numerous covalently bonded luminescent units.They have excellent light-harvesting and optical signal amplification capabilities owing to the transmission of excitation energy along their backbone.In recent years,CP materials have aroused research enthusiasm in the biosensors field because of their outstanding optoelectronic properties.This brief manuscript provides an overall review of recent progress achieved in CP-based systems for pathogen sensing.
基金Yong Science and Technology Talent of AAAS(QNYC-201911)。
文摘Fusarium crown rot,mainly caused by Fusarium pseudograminearum,is a destructive disease in wheat production.To establish a rapid and reliable detection method for F.peasudeograminearum,the specific PCR primer pair(Fpg-F1;R2)was designed based on the RPB sequence,and real-time fluorescence quantitative PCR(qPCR)was used to validate the efficiency of the primer.The results showed that the primer pair had high specificity and sensitivity of 100 pg of DNA.Furthermore,the qPCR system for early and rapid detection of F.peasudeograminearum had an amplification efficiency of 87.5%and correlation coefficient of 0.99,and the pathologic threshold of F.pseudograminearum in soil was determined by using this detection system.It was found that F.pseudograminearum could cause Fusarium crown rot when the DNA concentration of F.pseudograminearum in field soil exceeded 213 pg·g^(-1).Hence,the qPCR-based method we developed for F.pseudograminearum detection has the advantages of high specificity and sensitivity,and can be used for rapid and early detection of F.pseudograminearum even in field soils.
文摘In recent years,with the rapid development of molecular biology diagnostic technology,many new polymerase chain reaction(PCR)technologies with high specificity and good sensitivity have gradually been developed.While expanding the range of detection methods,these technologies inevitably have some disadvantages.Therefore,in clinical pathogen diagnosis,medical personnel should choose the detection method according to the detection purpose and pathogen characteristics.In this paper,the basic principle,application scope,advantages and disadvantages and development of various emerging PCR diagnostic techniques are respectively described in order to provide a theoretical reference for the selection of pathogenic biological diagnostic techniques in the clinical practice.
基金supported by grants from the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(No.2022-I2M-CoV19-006)the Science & Technology Fundamental Resources Investigation Program(No.2023FY100600)+1 种基金the Investigator Initiated Study in Collaboration with Sanofi(No.FLU00152)the Chongqing Science and Technology Bureau(Nos.CSTC2021jscx-gksb-N0005 and cstc2024ycjhbgzxm0224).
文摘To the Editor:Multi-pathogen detection has become increasingly important,given the increasing risk of circulating contagious pathogens due to global changes characterized by urbanization and habitat fragmentation.The coronavirus disease-2019(COVID-19)pandemic disrupted the epidemiology of some respiratory pathogens,possibly due to public health and social measures,as well as changes in healthcare-seeking behaviors and interactions between viruses.Multi-respiratory pathogen surveillance could be useful for adjusting disease prevention strategies and preventing potential outbreaks of existing and emerging respiratory infectious diseases.On the other hand,symptoms and disease severity differ among respiratory pathogen infections,[1]and coinfection might be associated with increased disease severity.[2]Some respiratory pathogens such as severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),influenza viruses(IFV),respiratory syncytial virus(RSV),and Streptococcus pneumoniae(S.pneumoniae)impose a high disease burden on populations but can be prevented by vaccines.Therefore,this study aimed to determine clinical manifestations and disease severity among patients testing positive for various pathogens to offer guidance on disease diagnosis,treatment,surveillance,and immunization strategies.
基金supported by Horizontal Project of Xinchang County Hospital of Chinese Medicine(2023‐HT‐394).
文摘Rare infectious diseases are infections that are uncommon,have a low incidence,and are caused by newly emerging pathogens,cross‐species or ectopic infections,or host immunodeficiencies.The detection and diagnosis of rare infections is one of the main reasons for misdiagnosis and missed diagnosis.Traditional detection techniques such as microbial cultivation and isolation or polymerase chain reaction often fail to meet the clinical demands for timeliness,accuracy,and sensitivity.Metagenomic nextgeneration sequencing involves pan-nucleic acid testing conducted directly using specimens to facilitate rapid identification of rare or unidentified pathogens.Despite the availability of various techniques,advanced methods in clinical practice are necessary to achieve timely clinical diagnosis of rare infections.In this review,we summarize the definition and clinical significance of rare infectious diseases as well as the current detection methods,limitations,and future research areas for their detection.
基金Supported by Guizhou Provincial Department of Agriculture and Rural Affairs Project(QNYZZZ[2017]No.12,GZSZCYJSTX-04)2025 Quality Supervision and Sampling Project of Normal Temperature Semen for Breeding Pigs(2025-1-10).
文摘[Objectives]This study was conducted to evaluate the immunization efficacy and infection status of classical swine fever(CSF),foot-and-mouth disease(FMD),porcine reproductive and respiratory syndrome(PRRS),pseudorabies(PR),and porcine circovirus type 2(PCV2)in large-scale pig farms.[Methods]Antibody and pathogen detection was performed on 56 serum samples collected in March 2025.[Results]The antibody qualification rates for CSF,FMD,and PRRS were 76.8%,73.2%,and 76.8%,respectively,all meeting the national standards.However,nursery pigs exhibited an immunity gap,indicating a need for timely booster vaccinations.No PRV gE antibodies or PCV2 antibodies were detected,reflecting the absence of vaccination against these diseases and suggesting significant effectiveness of comprehensive biosecurity measures.The low antibody qualification rate for PRRS in the nursery stage highlights the need for improved immunization management.[Conclusions]This study provides data support and practical insights for integrated disease prevention and control in large-scale pig farms.
基金supported by the National Natural Science Foundation of China(NSFC)(Nos.61935010,61975069,21905253,and 51973200)the China Postdoctoral Science Foundation(Nos.2018M640681 and 2019T120632)+5 种基金the Natural Science Foundation of Henan(No.202300410372)Key-Area Research and Development Program of Guangdong Province(No.2020B090922006)Guangdong Project of Science and Technology Grants(No.2018B030323017)Guangzhou science and technology project(Nos.201903010042 and 201904010294)Youth project of science and technology research program of Chongqing Education Commission of China(No.KJQN202001322)the Science and Technology Development Fund from Macao SAR(File Nos.0125/2018/A3 and 0071/2019/AMJ).
文摘The most widely used method of identification of microbial morphology and structure is microscopy,but it can be difficult to distinguish between pathogens with a similar appearance.Existing fluorescent staining methods require a combination of a variety of fluorescent materials to meet this demand.In this study,unique concentration-dependent fluorescent carbon dots(CDs)were synthesized for the identification and quantification of pathogens.The emission wavelength of the CDs could be tuned spanning the full visible region by virtue of aggregation-induced narrowing of bandgaps.This tunable emission wavelength of the specific concentration response to diverse microbes can be used to distinguish microorganisms with a similar appearance,even in a same genus.A hyperspectral microscopy system was demonstrated to distinguish Aspergillus flavus and A.fumigatus based on the results above.The identification accuracy of the two similar-looking pathogens can be close to 100%,and the relative proportions and spatial distributions can also be profiled from the mixture of the pathogens.This technique can provide a solution to the fast detection of microorganisms and is potentially applicable to a wide range of problems in areas such as healthcare,food preparation,biotechnology,and health emergency.
文摘Rapid,high-throughput,timely,multiplex diagnosis of respiratory-tract infections still relies on laboratory infrastructure,sequential assays,and trained personnel,thereby delaying targeted therapy and outbreak containment.In this study,a Fully Automated rotary microfluidic platform(FA-RMP)for high-throughput multiplex respiratory tract pathogens detection was presented.FA-RMP enables a true“sample-in,result-out”workflow through the integration of swab lysis,reagent partitioning,lyophilized reverse transcription loop-mediated isothermal amplification(RT-LAMP),and movingprobe fluorescence read-out,all encapsulated with a disposable microfluidic cartridge and paired with a 9 kg,fourchannel benchtop reader.The FA-RMP enables parallel processing of 16 independent reactions within 30 min,supporting simultaneous detection of up to 4 distinct clinical samples.Analytical validation using serially diluted Mycoplasma pneumoniae(MP)DNA established a limit of detection(LoD)of 50 copies μL^(-1) and a log-linear correlation between threshold time and template load(R^(2)=0.9528).Testing with eight non-target respiratory pathogens yielded no amplification,confirming high analytical specificity.FA-RMP successfully detected the clinical samples with influenza A,influenza B,and MP,further demonstrating its robust multiplex detection capability.By integrating automated sample preparation,multiplex isothermal amplification and quantitative detection into a portable,high-throughput system,the platform delivers laboratory-grade performance at the point of care,serving as a scalable tool for routine respiratory pathogens screening and rapid epidemic response.
基金Supported by Shandong Province Key Research and Development Plan(Major Scientific and Technological Innovation Project,2023CXGC010711)the National Key R&D Program of China(2021YFC2301102)the National Natural Science Foundation of China(82202593,U23A20106).
文摘Introduction:Fluorescent probe-based recombinase aided amplification(RAA)offers the advantages of rapidity and simplicity but is limited by the requirement for complex and lengthy probe design,restricting its widespread application.Methods:A novel EvaGreen dye-based RAA(EvaGreen-RAA)assay utilizing self-avoiding molecular recognition system(SAMRS)primers was developed for the detection of Pseudomonas fluorescens(PF)and Bacillus cereus(BC)in milk.Conventional RAA was used as a reference method.Sensitivity was evaluated using nucleic acids from recombinant plasmids and simulated milk specimens.Additionally,a dual EvaGreen-RAA assay was investigated for simultaneous detection of mixed BC and PF in simulated milk specimens.Results:The EvaGreen-RAA demonstrated superior sensitivity compared to conventional RAA,with detection limits of 1 copy/μL versus 10 copies/μL for both BC and PF plasmids,respectively.In simulated milk specimens,EvaGreen-RAA detected BC and PF at concentrations of 100 CFU/mL and 200 CFU/mL,respectively,compared to 400 CFU/mL and 600 CFU/mL for conventional RAA.The dual EvaGreen-RAA assay successfully detected mixed BC and PF in simulated milk specimens at concentrations of 200 CFU/mL for each pathogen.Conclusion:The EvaGreen-RAA assay demonstrated significant advantages in terms of simplicity and enhanced sensitivity compared to fluorescent probe-based RAA,offering a novel approach for developing multiplex pathogen detection systems using melting curve analysis.
基金European Commission(CZ.10.03.01/00/22-003/0000048)Fundacao para a Ciencia e a Tecnologia(PTDC/EEI-EEE/0415/2021),CICECO(UIDB/50011/2020,UIDP/50011/2020,LA/P/0006/2020)+1 种基金VSB-Technical University of Ostrava(SP2025/039)FCT/MCTES(UI/BD/153066/2022)。
文摘In the food production sector,quickly identifying potential hazards is crucial due to the resilience of many pathogens,which could lead to wasted production results and,more severely,epidemic outbreaks.E.coli monitoring is essential;however,traditional quality control methods in fish farming are often slow and intrusive,thus promoting an increase in fish stress and mortality rates.This paper presents an alternative method by utilizing a prototype inspired by polarized optical microscopy(POM),constructed with a Raspberry Pi microprocessor to assess pixel patterns and calculate analyte levels.
基金supported by grants from the National Natural Science Foundation of China(32372483,32302296,32272641)the Fundamental Research Funds for the Central Universities(GK202201017)。
文摘Some plants use intracellular immune receptors,mainly from the nucleotide-binding leucine-rich repeat receptors(NLRs)super-family,to detect pathogen effectors and restrict pathogen infection.NLRs are categorized as TIR-NLR(TNL)or CC-NLR(CNL)based on their N-terminal Tollinterleukin 1-like receptor(TIR)or coiled-coil(CC)domain.Additionally,there are helper(h)NLRs that aid in translating signals from pathogen-sensing NLRs into effector-triggered immunity responses.NLRs can directly detect effectors or indirectly monitor their host targets,leading to a robust immune response and programmed cell death,known as the hypersensitive response(HR).
基金supported by the UKRI NERC Fellowship(NE/R013349/2)the UKRI NERC N-WESP(NE/V010441/1)+1 种基金the NERC Innovative Sensing(NE/Z503538/1)the Leverhulme Trust Research Leadership Awards(RL-2022-041)。
文摘A sustainable early warning monitoring system can overcome key limitations of conventional approaches for managing infectious diseases and epidemics.It enables consistent monitoring of pathogens,including SARSCoV-2,in wastewater treatment plants(WWTPs).Wastewater-based epidemiology(WBE)analyses biological markers excreted in sewage to monitor community health.It has emerged as a powerful tool because it is cost-effective,non-invasive,and capable of detecting pathogen circulation within the population,even before clinical cases are reported[1].These features provide WBE a clear advantage over traditional clinical testing,particularly by enabling the early detection of new variants[2].
基金supported by the National Science Foundation(IOS-2207677 to Z.Q.F.)the National Natural Science Foundation of China(32260653 to C.L.)+1 种基金Guizhou Provincial General Project of the Science Foundation(ZK[2024]Key 011 to C.L.)the Scientific Research and Innovation Team of Guizhou University([2024]05 to C.L.).
文摘Sensory mechanisms play pivotal roles in plant defense against pathogen infection.To safeguard themselves,plants developed both cell surface-situated pattern recognition receptors(PRRs)to identify pathogen-associated molecular patterns(PAMPs)and intracellular nucleotidebinding leucine-rich repeat receptors(NLRs)to detect pathogen effectors when they face imminent threats from a wide range of pathogens(Boller and Felix,22009:Jones et al.,2024;Wu et al.,2014).
