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
Japanese encephalitis(JE)was first discovered in Japan in 1871;in 1924,a major outbreak occurred,with 6,000 JE cases reported and a mortality rate of approximately 60%[1,2].Later studies showed that JE is caused by th...Japanese encephalitis(JE)was first discovered in Japan in 1871;in 1924,a major outbreak occurred,with 6,000 JE cases reported and a mortality rate of approximately 60%[1,2].Later studies showed that JE is caused by the Japanese encephalitis virus(JEV),which is spread by mosquitoes.展开更多
Wuxiang virus(WUXV),a new species of Phlebovirus from sandfly specimens(identified as Phlebotomus chinensis)collected in Wuxiang County,Shanxi Province,China,belongs to the order Bunyavirales,family Phenuiviridae.Like...Wuxiang virus(WUXV),a new species of Phlebovirus from sandfly specimens(identified as Phlebotomus chinensis)collected in Wuxiang County,Shanxi Province,China,belongs to the order Bunyavirales,family Phenuiviridae.Like other Bunyavirales,WUXV contains a negative-sense tripartite RNA genome comprising a small(S),a medium(M),and a large(L)segment.The S segment encodes nucleocapsid protein(NP)and a nonstructural protein(NSP).The M segment encodes a glycoprotein precursor(Gp),which is cleaved into mature N and C glycoproteins.The L segment encodes an RNA-dependent RNA polymerase(RdRp).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 grants from the National Key Research and Development Program[2016YFD0500401](WH)the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control[2014SKLID103](LG)and[2015SKLID505](WH)。
文摘Japanese encephalitis(JE)was first discovered in Japan in 1871;in 1924,a major outbreak occurred,with 6,000 JE cases reported and a mortality rate of approximately 60%[1,2].Later studies showed that JE is caused by the Japanese encephalitis virus(JEV),which is spread by mosquitoes.
基金funded by a development grant from the State Key Laboratory of Infectious Disease Prevention and Control [2015SKLID505 to HW]the project of the Priority Academic Program Development of Jiangsu Higher Education Institutions [PAPD]the United States National Institutes of Health U01 [AI151810]
文摘Wuxiang virus(WUXV),a new species of Phlebovirus from sandfly specimens(identified as Phlebotomus chinensis)collected in Wuxiang County,Shanxi Province,China,belongs to the order Bunyavirales,family Phenuiviridae.Like other Bunyavirales,WUXV contains a negative-sense tripartite RNA genome comprising a small(S),a medium(M),and a large(L)segment.The S segment encodes nucleocapsid protein(NP)and a nonstructural protein(NSP).The M segment encodes a glycoprotein precursor(Gp),which is cleaved into mature N and C glycoproteins.The L segment encodes an RNA-dependent RNA polymerase(RdRp).
