Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laborat...Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing.Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing.Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing.Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.展开更多
The rapid spread of viral zoonoses can cause severe consequences,including huge economic loss,public health problems or even global crisis of society.Clinical detection technology plays a very important role in the pr...The rapid spread of viral zoonoses can cause severe consequences,including huge economic loss,public health problems or even global crisis of society.Clinical detection technology plays a very important role in the prevention and control of such zoonoses.The rapid and accurate detection of the pathogens of the diseases can directly lead to the early report and early successful control of the diseases.With the advantages of being easy to use,fast,portable,multiplexing and cost-effective,semiconductor biosensors are kinds of detection devices that play an important role in preventing epidemics,and thus have become one of the research hotspots.Here,we summarized the advances of semiconductor biosensors in viral zoonoses detection.By discussing the major principles and applications of each method for different pathogens,this review proposed the directions of designing semiconductor biosensors for clinical application and put forward perspectives in diagnostic of viral zoonoses.展开更多
Epstein-Barr virus(EBV)and cytomegalovirus(CMV),two of the most prevalent human herpesviruses,cause a wide spectrum of diseases and symptoms and are associated with serious health problem.In this study,we developed an...Epstein-Barr virus(EBV)and cytomegalovirus(CMV),two of the most prevalent human herpesviruses,cause a wide spectrum of diseases and symptoms and are associated with serious health problem.In this study,we developed an internal control reference recombinase-aided amplification(ICR-RAA)assay for the rapid detection of EBV and CMV within 30 min.The assay had a sensitivity of 5 and 1 copies/test for EBV and CMV,respectively,with no cross reaction with other pathogens.In comparison with those of the commercial quantitative polymerase chain reaction(qPCR),the sensitivity of the EBV and CMV ICR-RAAs using extracted DNA was 93.33%and 84.84%,respectively;the specificity was 98.75%and 100.00%,respectively;and the Kappa values were 0.930 and 0.892(P<0.05),respectively.In comparison with those of qPCR,the sensitivity of the EBV and CMV ICR-RAAs using the DNA by thermal lysis was 72.22%and 80.00%,respectively;the specificity was 100.00%。展开更多
West Nile virus(WNV)causes West Nile fever and West Nile encephalitis.Because infection by WNV creates serious public health problems,its simple,rapid,and visual detection is very important in clinical practice,especi...West Nile virus(WNV)causes West Nile fever and West Nile encephalitis.Because infection by WNV creates serious public health problems,its simple,rapid,and visual detection is very important in clinical practice,especially in resource-limited laboratories.We have developed a rapid,specific,and highly sensitive internally controlled reverse transcription recombinase-aided amplification(RTRAA)assay to detect WNV,using both real-timefluoresce nee and the lateral flow dipstick(LFD)at39.0°C for 30 min.The analytical sensitivity of theRT-RAA assay was 10 plasmid copies and 1.6 pfu perreacti on with real-time fluoresce nee,and 1,000plasmid copies per reaction with the LFD.No crossreactionwith other control viruses was observed.Compared with the RT-qPCR assay,the RT-RAA assaydemonstrated 100%sensitivity and 100%specificityfor WNV.展开更多
Parkinson’s disease(PD)is a typical degenerative disease,which is characterized by the most obvious symptoms of movement dysfunction,including shaking,rigidity,slowness of movement and difficulty in walking and gait....Parkinson’s disease(PD)is a typical degenerative disease,which is characterized by the most obvious symptoms of movement dysfunction,including shaking,rigidity,slowness of movement and difficulty in walking and gait.This disease can not be clearly identified through laboratory tests at present,thus application of high-throughput technique in studying展开更多
Objective This study explored the potentially modifiable factors for depression and major depressive disorder(MDD)from the MR-Base database and further evaluated the associations between drug targets with MDD.Methods ...Objective This study explored the potentially modifiable factors for depression and major depressive disorder(MDD)from the MR-Base database and further evaluated the associations between drug targets with MDD.Methods We analyzed two-sample of Mendelian randomization(2SMR)using genetic variant depression(n=113,154)and MDD(n=208,811)from Genome-Wide Association Studies(GWAS).Separate calculations were performed with modifiable risk factors from MR-Base for 1,001 genomes.The MR analysis was performed by screening drug targets with MDD in the DrugBank database to explore the therapeutic targets for MDD.Inverse variance weighted(IVW),fixed-effect inverse variance weighted(FE-IVW),MR-Egger,weighted median,and weighted mode were used for complementary calculation.Results The potential causal relationship between modifiable risk factors and depression contained 459 results for depression and 424 for MDD.Also,the associations between drug targets and MDD showed that SLC6A4,GRIN2A,GRIN2C,SCN10A,and IL1B expression are associated with an increased risk of depression.In contrast,ADRB1,CHRNA3,HTR3A,GSTP1,and GABRG2 genes are candidate protective factors against depression.Conclusion This study identified the risk factors causally associated with depression and MDD,and estimated 10 drug targets with significant impact on MDD,providing essential information for formulating strategies to prevent and treat depression.展开更多
Objective This study aimed to compare the current Essen rabies post-exposure immunization schedule(0-3-7-14-28)in China and the simple 4-dose schedule(0-3-7-14)newly recommended by the World Health Organization in ter...Objective This study aimed to compare the current Essen rabies post-exposure immunization schedule(0-3-7-14-28)in China and the simple 4-dose schedule(0-3-7-14)newly recommended by the World Health Organization in terms of their safety,efficacy,and protection.Methods Mice were vaccinated according to different immunization schedules,and blood was collected for detection of rabies virus neutralizing antibodies(RVNAs)on days 14,21,28,35,and 120after the first immunization.Additionally,different groups of mice were injected with lethal doses of the CVS-11 virus on day 0,subjected to different rabies immunization schedules,and assessed for morbidity and death status.In a clinical trial,185 rabies-exposed individuals were selected for post-exposure vaccination according to the Essen schedule,and blood was collected for RVNAs detection on days 28and 42 after the first immunization.Results A statistically significant difference in RVNAs between mice in the Essen and 0-3-7-14 schedule groups was observed on the 35th day(P<0.05).The groups 0-3-7-14,0-3-7-21,and 0-3-7-28 showed no statistically significant difference(P>0.05)in RVNAs levels at any time point.The post-exposure immune protective test showed that the survival rate of mice in the control group was 20%,whereas that in the immunization groups was 40%.In the clinical trial,the RVNAs positive conversion rates on days 28(14 days after 4 doses)and 42(14 days after 5 doses)were both 100%,and no significant difference in RVNAs levels was observed(P>0.05).Conclusion The simple 4-dose schedule can produce sufficient RVNAs levels,with no significant effect of a delayed fourth vaccine dose(14–28 d)on the immunization potential.展开更多
Objectives Hand,foot and mouth disease(HFMD)is a widespread infectious disease that causes a significant disease burden on society.To achieve early intervention and to prevent outbreaks of disease,we propose a novel w...Objectives Hand,foot and mouth disease(HFMD)is a widespread infectious disease that causes a significant disease burden on society.To achieve early intervention and to prevent outbreaks of disease,we propose a novel warning model that can accurately predict the incidence of HFMD.Methods We propose a spatial-temporal graph convolutional network(STGCN)that combines spatial factors for surrounding cities with historical incidence over a certain time period to predict the future occurrence of HFMD in Guangdong and Shandong between 2011 and 2019.The 2011-2018 data served as the training and verification set,while data from 2019 served as the prediction set.Six important parameters were selected and verified in this model and the deviation was displayed by the root mean square error and the mean absolute error.Results As the first application using a STGCN for disease forecasting,we succeeded in accurately predicting the incidence of HFMD over a 12-week period at the prefecture level,especially for cities of significant concern.Conclusions This model provides a novel approach for infectious disease prediction and may help health administrative departments implement effective control measures up to 3 months in advance,which may significantly reduce the morbidity associated with HFMD in the future.展开更多
Hand,foot,and mouth disease (HFMD) is a common contagious illness which occurs worldwide both sporadically and in epidemics.The disease mainly affects children and the typical symptoms,which may resolve spontaneously,...Hand,foot,and mouth disease (HFMD) is a common contagious illness which occurs worldwide both sporadically and in epidemics.The disease mainly affects children and the typical symptoms,which may resolve spontaneously,include mucocutaneous papulovesicular lesions on the hands,feet,mouth,and buttocks.In rare cases,however,the patients may also develop neurological complications such as neurogenic pulmonary edema,aseptic meningitis,and acute flaccid paralysis[1].The most common etiological agents of HFMD are human enterovirus 71 (EV-A71) and展开更多
We are thrilled to present this dedicated issue spotlighting enterovirus infections and health in Biosafety and Health.The articles presented in this issue span the broad spectrum of current research in this vital fie...We are thrilled to present this dedicated issue spotlighting enterovirus infections and health in Biosafety and Health.The articles presented in this issue span the broad spectrum of current research in this vital field,providing insights into the epidemiology,diagnostic procedures,and pathogenesis of enterovirus infections.Enteroviruses,a genus of RNA viruses encompassing polioviruses,coxsackieviruses,and echoviruses,play a crucial role in public health.展开更多
In 2022,a global outbreak of Mpox(formerly monkeypox)occurred in various countries across Europe and America and rapidly spread to more than 100 countries and regions.The World Health Organization declared the outbrea...In 2022,a global outbreak of Mpox(formerly monkeypox)occurred in various countries across Europe and America and rapidly spread to more than 100 countries and regions.The World Health Organization declared the outbreak to be a public health emergency of international concern due to the rapid spread of the Mpox virus.Consequently,nations intensified their efforts to explore treatment strategies aimed at combating the infection and its dissemination.Nevertheless,the available therapeutic options for Mpox virus infection remain limited.So far,only a few numbers of antiviral compounds have been approved by regulatory authorities.Given the high mutability of the Mpox virus,certain mutant strains have shown resistance to existing pharmaceutical interventions.This highlights the urgent need to develop novel antiviral drugs that can combat both drug resistance and the potential threat of bioterrorism.Currently,there is a lack of comprehensive literature on the pathophysiology and treatment of Mpox.To address this issue,we conducted a review covering the physiological and pathological processes of Mpox infection,summarizing the latest progress of anti-Mpox drugs.Our analysis encompasses approved drugs currently employed in clinical settings,as well as newly identified small-molecule compounds and antibody drugs displaying potential antiviral efficacy against Mpox.Furthermore,we have gained valuable insights from the process of Mpox drug development,including strategies for repurposing drugs,the discovery of drug targets driven by artificial intelligence,and preclinical drug development.The purpose of this review is to provide readers with a comprehensive overview of the current knowledge on Mpox.展开更多
Objective This study was conducted to investigate the viral and bacterial etiology and epidemiology of patients with acute febrile respiratory syndrome(AFRS) in Qinghai using a commercial routine multiplex-ligation-nu...Objective This study was conducted to investigate the viral and bacterial etiology and epidemiology of patients with acute febrile respiratory syndrome(AFRS) in Qinghai using a commercial routine multiplex-ligation-nucleic acid amplification test(NAT)-based assay. Methods A total of 445 nasopharyngeal swabs specimens from patients with AFRS were analyzed using the RespiFinderSmart22 kit(PathoFinder BV, Netherlands) and the LightCycler 480 real-time PCR system. Results Among the 225(225/445, 51%) positive specimens, 329 positive pathogens were detected, including 298(90.58%) viruses and 31(9%) bacteria. The most commonly detected pathogens were influenza virus(IFV;37.39%;123/329), adenovirus(AdV;17.02%;56/329), human coronaviruses(HCoVs;10.94%;36/329), rhinovirus/enterovirus(RV/EV;10.03%;33/329), parainfluenza viruses(PIVs;8.51%;28/329), and Mycoplasma pneumoniae(M. pneu;8.51%;28/329), respectively. Among the co-infected cases(17.53%;78/445), IFV/AdV and IFV/M. pneu were the most common co-infections. Most of the respiratory viruses were detected in summer and fall. Conclusion In our study, IFV-A was the most common respiratory pathogen among 22 detected pathogens, followed by AdV, HCoV, RV/EV, PIV, and M. pneu. Bacteria appeared less frequently than viruses, and co-infection was the most common phenomenon among viral pathogens. Pathogens were distributed among different age groups and respiratory viruses were generally active in July, September, and November. Enhanced surveillance and early detection can be useful in the diagnosis, treatment, and prevention of AFRS, as well as for guiding the development of appropriate public health strategies.展开更多
Objective To detect Japanese encephalitis virus(JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction(RT-PCR) detection system was developed.Methods By...Objective To detect Japanese encephalitis virus(JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction(RT-PCR) detection system was developed.Methods By aligning the full-length sequences of JEV(G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.Results With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/μL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.Conclusion A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes.展开更多
Objective To provide a feasible and cost-effective next-generation sequencing(NGS) method for accurate identification of viral pathogens in clinical specimens, because enormous limitations impede the clinical use of c...Objective To provide a feasible and cost-effective next-generation sequencing(NGS) method for accurate identification of viral pathogens in clinical specimens, because enormous limitations impede the clinical use of common NGS, such as high cost, complicated procedures, tremendous data analysis, and high background noise in clinical samples. Methods Viruses from cell culture materials or clinical specimens were identified following an improved NGS procedure: reduction of background noise by sample preprocessing, viral enrichment by barcoded oligonucleotide(random hexamer or non-ribosomal hexanucleotide) primer-based amplification, fragmentation-free library construction and sequencing of one-tube mixtures, as well as rapid data analysis using an in-house pipeline. Results NGS data demonstrated that both barcoded primer sets were useful to simultaneously capture multiple viral pathogens in cell culture materials or clinical specimens and verified that hexanucleotide primers captured as many viral sequences as hexamers did. Moreover, direct testing of clinical specimens using this improved hexanucleotide primer-based NGS approach provided further detailed genotypes of enteroviruses causing hand, foot, and mouth disease(HFMD) and identified other potential viruses or differentiated misdiagnosis events. Conclusion The improved barcoded oligonucleotide primer-based NGS approach is simplified, time saving, cost effective, and appropriate for direct identification of viral pathogens in clinical practice.展开更多
Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of path...Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection,inflammation,and CHD,to provide a basis for the prevention,evaluation,and treatment of the disease.Methods In total,192 patients with CHD were divided into three groups:latent CHD,angina pectoris,and myocardial infarction.HCMV-IgM and-IgG antibodies were assessed using ELISA;CD14+CD16+monocytes were counted using a five-type automated hematology analyzer;mononuclear cells were assessed using fluorescence-activated cell sorting;and an automatic biochemical analyzer was used to measure the levels of triglyceride,cholesterol,high-and low-density lipoprotein cholesterols,lipoprotein,hs-CRp and Hcy.Results The positive rates of HCMV-IgM and-IgG were significantly higher in the CHD groups than in the control group.HCMV infection affects lipid metabolism to promote immune and inflammatory responses.Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD.The expression of CD14+CD16+mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection.Thus,HCMV antibody as well as peripheral blood CD14+CD16+mononuclear cells can be used to monitor the occurrence and development of CHD.展开更多
Objective Newly identified human rhinovirus C(HRV-C) and human bocavirus(HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines...Objective Newly identified human rhinovirus C(HRV-C) and human bocavirus(HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses. Methods A platform for culturing human airway epithelia in a three-dimensional(3 D) pattern using Matrigel as scaffold was developed. The features of 3 D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3 D cells at designated time points were quantitated by real-time polymerase chain reaction(PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA. Results Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3 D-cultured human airway epithelial(HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3 D culture system. Conclusion Our data provide a preliminary indication that the 3 D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.展开更多
Objective Hepatitis Delt a Virus(HDV) antigen is widely used as a capture antigen in ELISAs for the identification of HDV infection; large amounts of recombinant HDV antigen with active antigenicity are required for t...Objective Hepatitis Delt a Virus(HDV) antigen is widely used as a capture antigen in ELISAs for the identification of HDV infection; large amounts of recombinant HDV antigen with active antigenicity are required for this purpose. Methods Reconstruct the gene of HDV antigen based on the bias code of Escherichia coli, the recombinant protein expresses by high-density fermentation with fed-batch feeding strategy, and purify by immobilized metal chromatography. The sensitivity and specificity of this antigen detect by ELISA method. Results The expression of HDV antigen can reach 20% of the total cell mass in the soluble form. The recombinant HDV antigen can be conveniently purified(98%) by immobilized metal ion affinity chromatography(IMAC) using the interaction between a His-tag and nickel ions. Production of recombinant HDV antigen can reach 0.5 g/L under conditions of high-density cell fermentation. Applied to the diagnostic ELISA method, the recombinant HDV antigen shows excellent sensitivity(97% for IgM and 100% for IgG) and specificity(100% for IgG and IgM) for the detection of anti-HDV antibodies. Conclusion Expression and purification the recombinant HDV antigen as a candidate protein for application in a diagnostic ELISA for HDV infection. Large-scale production of the protein can be achieved using the high-density fermentation strategy.展开更多
Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor (VEGF) in sera of population and the patients with hepatocellular carcinoma (HCC). Methods Full length and two truncated ...Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor (VEGF) in sera of population and the patients with hepatocellular carcinoma (HCC). Methods Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-hVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms. The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165. After purification of antibodies with chromatograph of Protein G, a sandwich ELISA technique was established. Serum VEGF levels were evaluated in 229 adults and 291 HCC patients. Results SDS-PAGE displayed that the molecular weights of the expressed full length (His-Trx-VEGF1-165), N-terminal (His-Trx-VEGF1-100) and C-terminal (GST-VEGF100-165) human VEGF fusion proteins were about 38KD, 31KD, and 33KD, respectively. Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins. Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL, with the squared correlation coefficient R2=0.991. Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults revealed that the average VEGF level in HCC patients was higher than that in healthy controls, with a statically significant difference. Conclusion The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for screening metastasis and recurrence of HCC using serum VEGF as an index.展开更多
日本脑炎(JE ) 是一个严重公共健康问题。这研究被承担更好在中国理解在 JE 分发和环境因素之间的关系。从 2005 ～ 2010 的 JE 数据从国家应具报的疾病报告系统被检索。ArcGIS，遥感技术，和 R 软件被用来展出并且探索在 JE 分发和环...日本脑炎(JE ) 是一个严重公共健康问题。这研究被承担更好在中国理解在 JE 分发和环境因素之间的关系。从 2005 ～ 2010 的 JE 数据从国家应具报的疾病报告系统被检索。ArcGIS，遥感技术，和 R 软件被用来展出并且探索在 JE 分发和环境因素之间的关系。我们的结果显示 JE 案例主要与年度降水在温暖适度、亚热带、热带的地区被集中 > 400 公里；阔叶常绿树森林，灌木，稻地，灌溉土地， dryland，常绿树具球果的森林，和 shrubland 是为五是的 JE 出现，和前者的风险因素为有高 JE 的县的风险因素发生。这些调查结果将通知有效分配象集中的种痘那样的有限健康资源，监视并且与高环境的风险因素在区域训练。展开更多
基金supported by the National Key Research and Development Program(grant number:2022YFC2305304).
文摘Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing.Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing.Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing.Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.
基金supported by the National Key Research and Development Program of China(2022YFC2602100)supported by National key research and development program(2021YFC2600602)。
文摘The rapid spread of viral zoonoses can cause severe consequences,including huge economic loss,public health problems or even global crisis of society.Clinical detection technology plays a very important role in the prevention and control of such zoonoses.The rapid and accurate detection of the pathogens of the diseases can directly lead to the early report and early successful control of the diseases.With the advantages of being easy to use,fast,portable,multiplexing and cost-effective,semiconductor biosensors are kinds of detection devices that play an important role in preventing epidemics,and thus have become one of the research hotspots.Here,we summarized the advances of semiconductor biosensors in viral zoonoses detection.By discussing the major principles and applications of each method for different pathogens,this review proposed the directions of designing semiconductor biosensors for clinical application and put forward perspectives in diagnostic of viral zoonoses.
基金the China Mega-Projects for Infectious Disease[grant no.2018ZX10711001,2017ZX10104001 and 2018ZX10713-002]the National Institute for Viral Disease Control and Prevention[grant no.2019HYDQNJJ03].
文摘Epstein-Barr virus(EBV)and cytomegalovirus(CMV),two of the most prevalent human herpesviruses,cause a wide spectrum of diseases and symptoms and are associated with serious health problem.In this study,we developed an internal control reference recombinase-aided amplification(ICR-RAA)assay for the rapid detection of EBV and CMV within 30 min.The assay had a sensitivity of 5 and 1 copies/test for EBV and CMV,respectively,with no cross reaction with other pathogens.In comparison with those of the commercial quantitative polymerase chain reaction(qPCR),the sensitivity of the EBV and CMV ICR-RAAs using extracted DNA was 93.33%and 84.84%,respectively;the specificity was 98.75%and 100.00%,respectively;and the Kappa values were 0.930 and 0.892(P<0.05),respectively.In comparison with those of qPCR,the sensitivity of the EBV and CMV ICR-RAAs using the DNA by thermal lysis was 72.22%and 80.00%,respectively;the specificity was 100.00%。
基金supported by grants from the China Mega-Projects for Infectious Disease [2017ZX10302301-004-002,2018ZX10711001,2017ZX10104001,2018ZX10713-002]IVDC [2019HYDQNJJ03]
文摘West Nile virus(WNV)causes West Nile fever and West Nile encephalitis.Because infection by WNV creates serious public health problems,its simple,rapid,and visual detection is very important in clinical practice,especially in resource-limited laboratories.We have developed a rapid,specific,and highly sensitive internally controlled reverse transcription recombinase-aided amplification(RTRAA)assay to detect WNV,using both real-timefluoresce nee and the lateral flow dipstick(LFD)at39.0°C for 30 min.The analytical sensitivity of theRT-RAA assay was 10 plasmid copies and 1.6 pfu perreacti on with real-time fluoresce nee,and 1,000plasmid copies per reaction with the LFD.No crossreactionwith other control viruses was observed.Compared with the RT-qPCR assay,the RT-RAA assaydemonstrated 100%sensitivity and 100%specificityfor WNV.
基金supported by the National Natural Science Foundation of China(81101302,31270185)SKLID Development Grant(2014,SKLID201)
文摘Parkinson’s disease(PD)is a typical degenerative disease,which is characterized by the most obvious symptoms of movement dysfunction,including shaking,rigidity,slowness of movement and difficulty in walking and gait.This disease can not be clearly identified through laboratory tests at present,thus application of high-throughput technique in studying
基金supported by Natural Science Foundation of Shandong ProvinceChina[ZR2022MH115]the National Natural Science Foundation of China[81301479,82202593]。
文摘Objective This study explored the potentially modifiable factors for depression and major depressive disorder(MDD)from the MR-Base database and further evaluated the associations between drug targets with MDD.Methods We analyzed two-sample of Mendelian randomization(2SMR)using genetic variant depression(n=113,154)and MDD(n=208,811)from Genome-Wide Association Studies(GWAS).Separate calculations were performed with modifiable risk factors from MR-Base for 1,001 genomes.The MR analysis was performed by screening drug targets with MDD in the DrugBank database to explore the therapeutic targets for MDD.Inverse variance weighted(IVW),fixed-effect inverse variance weighted(FE-IVW),MR-Egger,weighted median,and weighted mode were used for complementary calculation.Results The potential causal relationship between modifiable risk factors and depression contained 459 results for depression and 424 for MDD.Also,the associations between drug targets and MDD showed that SLC6A4,GRIN2A,GRIN2C,SCN10A,and IL1B expression are associated with an increased risk of depression.In contrast,ADRB1,CHRNA3,HTR3A,GSTP1,and GABRG2 genes are candidate protective factors against depression.Conclusion This study identified the risk factors causally associated with depression and MDD,and estimated 10 drug targets with significant impact on MDD,providing essential information for formulating strategies to prevent and treat depression.
文摘Objective This study aimed to compare the current Essen rabies post-exposure immunization schedule(0-3-7-14-28)in China and the simple 4-dose schedule(0-3-7-14)newly recommended by the World Health Organization in terms of their safety,efficacy,and protection.Methods Mice were vaccinated according to different immunization schedules,and blood was collected for detection of rabies virus neutralizing antibodies(RVNAs)on days 14,21,28,35,and 120after the first immunization.Additionally,different groups of mice were injected with lethal doses of the CVS-11 virus on day 0,subjected to different rabies immunization schedules,and assessed for morbidity and death status.In a clinical trial,185 rabies-exposed individuals were selected for post-exposure vaccination according to the Essen schedule,and blood was collected for RVNAs detection on days 28and 42 after the first immunization.Results A statistically significant difference in RVNAs between mice in the Essen and 0-3-7-14 schedule groups was observed on the 35th day(P<0.05).The groups 0-3-7-14,0-3-7-21,and 0-3-7-28 showed no statistically significant difference(P>0.05)in RVNAs levels at any time point.The post-exposure immune protective test showed that the survival rate of mice in the control group was 20%,whereas that in the immunization groups was 40%.In the clinical trial,the RVNAs positive conversion rates on days 28(14 days after 4 doses)and 42(14 days after 5 doses)were both 100%,and no significant difference in RVNAs levels was observed(P>0.05).Conclusion The simple 4-dose schedule can produce sufficient RVNAs levels,with no significant effect of a delayed fourth vaccine dose(14–28 d)on the immunization potential.
基金supported by grants from the Key Technologies Research and Development Program from the Ministry of Science and Technology[grant number:ZDZX-2018ZX102001002-003-003]the Beijing Natural Science Foundation[project number:L192014]
文摘Objectives Hand,foot and mouth disease(HFMD)is a widespread infectious disease that causes a significant disease burden on society.To achieve early intervention and to prevent outbreaks of disease,we propose a novel warning model that can accurately predict the incidence of HFMD.Methods We propose a spatial-temporal graph convolutional network(STGCN)that combines spatial factors for surrounding cities with historical incidence over a certain time period to predict the future occurrence of HFMD in Guangdong and Shandong between 2011 and 2019.The 2011-2018 data served as the training and verification set,while data from 2019 served as the prediction set.Six important parameters were selected and verified in this model and the deviation was displayed by the root mean square error and the mean absolute error.Results As the first application using a STGCN for disease forecasting,we succeeded in accurately predicting the incidence of HFMD over a 12-week period at the prefecture level,especially for cities of significant concern.Conclusions This model provides a novel approach for infectious disease prediction and may help health administrative departments implement effective control measures up to 3 months in advance,which may significantly reduce the morbidity associated with HFMD in the future.
基金supported by National Foundation of China (project No.2013ZX10004-202)National Basic Research Program of China (973 Program,2011CB504902)National Natural Science Foundation of China (project Nos.30900063,81101303,81373049)
文摘Hand,foot,and mouth disease (HFMD) is a common contagious illness which occurs worldwide both sporadically and in epidemics.The disease mainly affects children and the typical symptoms,which may resolve spontaneously,include mucocutaneous papulovesicular lesions on the hands,feet,mouth,and buttocks.In rare cases,however,the patients may also develop neurological complications such as neurogenic pulmonary edema,aseptic meningitis,and acute flaccid paralysis[1].The most common etiological agents of HFMD are human enterovirus 71 (EV-A71) and
文摘We are thrilled to present this dedicated issue spotlighting enterovirus infections and health in Biosafety and Health.The articles presented in this issue span the broad spectrum of current research in this vital field,providing insights into the epidemiology,diagnostic procedures,and pathogenesis of enterovirus infections.Enteroviruses,a genus of RNA viruses encompassing polioviruses,coxsackieviruses,and echoviruses,play a crucial role in public health.
基金supported by National Natural Science Foundation of China(82002192)Natural Science Foundation of Hubei Province(2022CFB539,2022CFD107)+2 种基金Young and middle-aged Talents Project of Hubei Provincial Education Department(Q20222605)Scientific Research Ability Cultivation Fund of Hubei University of Arts and Science(2021KPGJ06)Science and Technology Plan(in the field of Medical and health care)of Xiangyang(2022YL05B,2022YL12A).
文摘In 2022,a global outbreak of Mpox(formerly monkeypox)occurred in various countries across Europe and America and rapidly spread to more than 100 countries and regions.The World Health Organization declared the outbreak to be a public health emergency of international concern due to the rapid spread of the Mpox virus.Consequently,nations intensified their efforts to explore treatment strategies aimed at combating the infection and its dissemination.Nevertheless,the available therapeutic options for Mpox virus infection remain limited.So far,only a few numbers of antiviral compounds have been approved by regulatory authorities.Given the high mutability of the Mpox virus,certain mutant strains have shown resistance to existing pharmaceutical interventions.This highlights the urgent need to develop novel antiviral drugs that can combat both drug resistance and the potential threat of bioterrorism.Currently,there is a lack of comprehensive literature on the pathophysiology and treatment of Mpox.To address this issue,we conducted a review covering the physiological and pathological processes of Mpox infection,summarizing the latest progress of anti-Mpox drugs.Our analysis encompasses approved drugs currently employed in clinical settings,as well as newly identified small-molecule compounds and antibody drugs displaying potential antiviral efficacy against Mpox.Furthermore,we have gained valuable insights from the process of Mpox drug development,including strategies for repurposing drugs,the discovery of drug targets driven by artificial intelligence,and preclinical drug development.The purpose of this review is to provide readers with a comprehensive overview of the current knowledge on Mpox.
基金supported by State Major Project of Infections Disease Control and Prevention of China [2017ZX10104001-002-003 and 2014ZX10004001-002]the National Key Research and Development Program of China [2016YFD0500301 and 2016YFC1200200]
文摘Objective This study was conducted to investigate the viral and bacterial etiology and epidemiology of patients with acute febrile respiratory syndrome(AFRS) in Qinghai using a commercial routine multiplex-ligation-nucleic acid amplification test(NAT)-based assay. Methods A total of 445 nasopharyngeal swabs specimens from patients with AFRS were analyzed using the RespiFinderSmart22 kit(PathoFinder BV, Netherlands) and the LightCycler 480 real-time PCR system. Results Among the 225(225/445, 51%) positive specimens, 329 positive pathogens were detected, including 298(90.58%) viruses and 31(9%) bacteria. The most commonly detected pathogens were influenza virus(IFV;37.39%;123/329), adenovirus(AdV;17.02%;56/329), human coronaviruses(HCoVs;10.94%;36/329), rhinovirus/enterovirus(RV/EV;10.03%;33/329), parainfluenza viruses(PIVs;8.51%;28/329), and Mycoplasma pneumoniae(M. pneu;8.51%;28/329), respectively. Among the co-infected cases(17.53%;78/445), IFV/AdV and IFV/M. pneu were the most common co-infections. Most of the respiratory viruses were detected in summer and fall. Conclusion In our study, IFV-A was the most common respiratory pathogen among 22 detected pathogens, followed by AdV, HCoV, RV/EV, PIV, and M. pneu. Bacteria appeared less frequently than viruses, and co-infection was the most common phenomenon among viral pathogens. Pathogens were distributed among different age groups and respiratory viruses were generally active in July, September, and November. Enhanced surveillance and early detection can be useful in the diagnosis, treatment, and prevention of AFRS, as well as for guiding the development of appropriate public health strategies.
基金supported by grants from the National Key Research and Development Program[2016YFD0500401]Development Grant of State Key Laboratory of Infectious Disease Prevention and Control[2015SKLID505,2014SKLID03]
文摘Objective To detect Japanese encephalitis virus(JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction(RT-PCR) detection system was developed.Methods By aligning the full-length sequences of JEV(G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.Results With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/μL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.Conclusion A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes.
基金supported by the China Mega-Project for Infectious Disease(2016ZX10004-101,2016ZX10004-215)Beijing Municipal Science&Technology Commission Project(D151100002115003)Guangzhou Municipal Science&Technology Commission Project(2015B2150820)
文摘Objective To provide a feasible and cost-effective next-generation sequencing(NGS) method for accurate identification of viral pathogens in clinical specimens, because enormous limitations impede the clinical use of common NGS, such as high cost, complicated procedures, tremendous data analysis, and high background noise in clinical samples. Methods Viruses from cell culture materials or clinical specimens were identified following an improved NGS procedure: reduction of background noise by sample preprocessing, viral enrichment by barcoded oligonucleotide(random hexamer or non-ribosomal hexanucleotide) primer-based amplification, fragmentation-free library construction and sequencing of one-tube mixtures, as well as rapid data analysis using an in-house pipeline. Results NGS data demonstrated that both barcoded primer sets were useful to simultaneously capture multiple viral pathogens in cell culture materials or clinical specimens and verified that hexanucleotide primers captured as many viral sequences as hexamers did. Moreover, direct testing of clinical specimens using this improved hexanucleotide primer-based NGS approach provided further detailed genotypes of enteroviruses causing hand, foot, and mouth disease(HFMD) and identified other potential viruses or differentiated misdiagnosis events. Conclusion The improved barcoded oligonucleotide primer-based NGS approach is simplified, time saving, cost effective, and appropriate for direct identification of viral pathogens in clinical practice.
基金Funded by the National Natural Science Foundation of China[81471048]the Natural Science Foundation of Shandong Province[ZR2019MC059]Shandong Province Government-Sponsored Overseas Study Project.&These authors contributed equally to this work.
文摘Objective To investigate the relationship between human cytomegalovirus(HCMV)infection and peripheral blood CD14+CD16+monocytes in the pathogenesis of coronary heart disease(CHD),and to elucidate the mechanism of pathogenesis in CHD by analyzing the correlation between infection,inflammation,and CHD,to provide a basis for the prevention,evaluation,and treatment of the disease.Methods In total,192 patients with CHD were divided into three groups:latent CHD,angina pectoris,and myocardial infarction.HCMV-IgM and-IgG antibodies were assessed using ELISA;CD14+CD16+monocytes were counted using a five-type automated hematology analyzer;mononuclear cells were assessed using fluorescence-activated cell sorting;and an automatic biochemical analyzer was used to measure the levels of triglyceride,cholesterol,high-and low-density lipoprotein cholesterols,lipoprotein,hs-CRp and Hcy.Results The positive rates of HCMV-IgM and-IgG were significantly higher in the CHD groups than in the control group.HCMV infection affects lipid metabolism to promote immune and inflammatory responses.Conclusion HCMV infection has a specific correlation with the occurrence and development of CHD.The expression of CD14+CD16+mononuclear cells in the CHD group was increased accordingly and correlated with acute HCMV infection.Thus,HCMV antibody as well as peripheral blood CD14+CD16+mononuclear cells can be used to monitor the occurrence and development of CHD.
基金supported by grants from the Major Project Specialized for Infectious Diseases of the Chinese Health and Family Planning Commission[2014ZX10004002-004-002,2014ZX10004002-004-001]Young Talent Scholar Plan of Higher School in Hebei Province[BJ2017008]
文摘Objective Newly identified human rhinovirus C(HRV-C) and human bocavirus(HBoV) cannot propagate in vitro in traditional cell culture models; thus obtaining knowledge about these viruses and developing related vaccines are difficult. Therefore, it is necessary to develop a novel platform for the propagation of these types of viruses. Methods A platform for culturing human airway epithelia in a three-dimensional(3 D) pattern using Matrigel as scaffold was developed. The features of 3 D culture were identified by immunochemical staining and transmission electron microscopy. Nucleic acid levels of HRV-C and HBoV in 3 D cells at designated time points were quantitated by real-time polymerase chain reaction(PCR). Levels of cytokines, whose secretion was induced by the viruses, were measured by ELISA. Results Properties of bronchial-like tissues, such as the expression of biomarkers CK5, ZO-1, and PCK, and the development of cilium-like protuberances indicative of the human respiration tract, were observed in 3 D-cultured human airway epithelial(HAE) cultures, but not in monolayer-cultured cells. Nucleic acid levels of HRV-C and HBoV and levels of virus-induced cytokines were also measured using the 3 D culture system. Conclusion Our data provide a preliminary indication that the 3 D culture model of primary epithelia using a Matrigel scaffold in vitro can be used to propagate HRV-C and HBoV.
文摘Objective Hepatitis Delt a Virus(HDV) antigen is widely used as a capture antigen in ELISAs for the identification of HDV infection; large amounts of recombinant HDV antigen with active antigenicity are required for this purpose. Methods Reconstruct the gene of HDV antigen based on the bias code of Escherichia coli, the recombinant protein expresses by high-density fermentation with fed-batch feeding strategy, and purify by immobilized metal chromatography. The sensitivity and specificity of this antigen detect by ELISA method. Results The expression of HDV antigen can reach 20% of the total cell mass in the soluble form. The recombinant HDV antigen can be conveniently purified(98%) by immobilized metal ion affinity chromatography(IMAC) using the interaction between a His-tag and nickel ions. Production of recombinant HDV antigen can reach 0.5 g/L under conditions of high-density cell fermentation. Applied to the diagnostic ELISA method, the recombinant HDV antigen shows excellent sensitivity(97% for IgM and 100% for IgG) and specificity(100% for IgG and IgM) for the detection of anti-HDV antibodies. Conclusion Expression and purification the recombinant HDV antigen as a candidate protein for application in a diagnostic ELISA for HDV infection. Large-scale production of the protein can be achieved using the high-density fermentation strategy.
基金National High-tech Development Project (863 Project) (No. 2002AA215021)
文摘Objective To establish a sandwich ELISA method for detecting vascular endothelial growth factor (VEGF) in sera of population and the patients with hepatocellular carcinoma (HCC). Methods Full length and two truncated human VEGF cDNA sequences were amplified from a commercial plasmid pBLAST49-hVEGF by PCR and inserted into the prokaryotic-expression plasmid pET-32a or pGEX-2T. Various VEGF proteins were expressed and purified from E. coli in His-Trx or GST fusion forms. The specific VEGF antibodies were elicited in experimental rabbits and mice by immunization of the full length VEGF fusion protein His-Trx-VEGF1-165. After purification of antibodies with chromatograph of Protein G, a sandwich ELISA technique was established. Serum VEGF levels were evaluated in 229 adults and 291 HCC patients. Results SDS-PAGE displayed that the molecular weights of the expressed full length (His-Trx-VEGF1-165), N-terminal (His-Trx-VEGF1-100) and C-terminal (GST-VEGF100-165) human VEGF fusion proteins were about 38KD, 31KD, and 33KD, respectively. Western blots confirmed that the prepared antisera were able to recognize both prokaryoticly and eukaryoticly expressed recombinant VEGF proteins. Assays of serially diluted His-Trx-VEGF1-100 by the established sandwich ELISA method showed that the linear range of the standard curve was 0.625-320 ng/mL, with the squared correlation coefficient R2=0.991. Screening of a serum panel containing 291 serum samples of HCC patients and 229 health adults revealed that the average VEGF level in HCC patients was higher than that in healthy controls, with a statically significant difference. Conclusion The established sandwich ELISA reflects the level of serum VEGF and provide scientific basis for screening metastasis and recurrence of HCC using serum VEGF as an index.
基金supported by the National Key Research and Development Program[2016YFD0500401]National Science and Technology Support Program[2014BAI13B05]the Development Grant of State Key Laboratory for Infectious Disease Prevention and Control[2015SKLID505]
文摘日本脑炎(JE ) 是一个严重公共健康问题。这研究被承担更好在中国理解在 JE 分发和环境因素之间的关系。从 2005 ～ 2010 的 JE 数据从国家应具报的疾病报告系统被检索。ArcGIS，遥感技术，和 R 软件被用来展出并且探索在 JE 分发和环境因素之间的关系。我们的结果显示 JE 案例主要与年度降水在温暖适度、亚热带、热带的地区被集中 > 400 公里；阔叶常绿树森林，灌木，稻地，灌溉土地， dryland，常绿树具球果的森林，和 shrubland 是为五是的 JE 出现，和前者的风险因素为有高 JE 的县的风险因素发生。这些调查结果将通知有效分配象集中的种痘那样的有限健康资源，监视并且与高环境的风险因素在区域训练。
