The Getah virus(GETV),a mosquito-borne RNA virus,is widely distributed in Oceania and Asia.GETV is not the only pathogenic to horses,pigs,cattle,foxes and boars,but it can also cause fever in humans.Since its first re...The Getah virus(GETV),a mosquito-borne RNA virus,is widely distributed in Oceania and Asia.GETV is not the only pathogenic to horses,pigs,cattle,foxes and boars,but it can also cause fever in humans.Since its first reported case in Chinese mainland in 2017,the number of GETV-affected provinces has increased to seventeen till now.Therefore,we performed an epidemiologic investigation of GETV in the Xinjiang region,located in northwestern China,during the period of 2017-2020.ELISA was used to analyze 3299 serum samples collected from thoroughbred horse,local horse,sheep,goat,cattle,and pigs,with thoroughbred horse(74.8%),local horse(67.3%),goat(11.7%),sheep(10.0%),cattle(25.1%)and pigs(51.1%)being positive for anti-GETV antibodies.Interestingly,the neutralizing antibody titer in horses was much higher than in other species.Four samples from horses and pigs were positive for GETV according to RT-PCR.Furthermore,from the serum of a local horse,we isolated GETV which was designated as strain XJ-2019-07,and determined its complete genome sequence.From the phylogenetic relationships,it belongs to the Group III lineage.This is the first evidence of GETV associated to domestic animals in Xinjiang.Overall,GETV is prevalent in Xinjiang and probably has been for several years.Since no vaccine against GETV is available in China,detection and monitoring strategies should be improved in horses and pigs,especially imported and farmed,in order to prevent economic losses.展开更多
Getah virus(GETV)is a mosquito-borne virus of the genus Alphavirus in the family Togaviridae and,in recent years,it has caused several outbreaks in animals.The molecular basis for GETV pathogenicity is not well unders...Getah virus(GETV)is a mosquito-borne virus of the genus Alphavirus in the family Togaviridae and,in recent years,it has caused several outbreaks in animals.The molecular basis for GETV pathogenicity is not well understood.Therefore,a reverse genetic system of GETV is needed to produce genetically modified viruses for the study of the viral replication and its pathogenic mechanism.Here,we generated a CMV-driven infectious cDNA clone based on a previously isolated GETV strain,GX201808(pGETV-GX).Transfection of pGETV-GX into BHK-21 cells resulted in the recovery of a recombinant virus(rGETV-GX)which showed similar growth characteristics to its parental virus.Then three-day-old mice were experimentally infected with either the parental or recombinant virus.The recombinant virus showed milder pathogenicity than the parental virus in the mice.Based on the established CMV-driven cDNA clone,subgenomic promoter and two restriction enzyme sites(BamHI and EcoRI)were introduced into the region between E1 protein and 3’UTR.Then the green fluorescent protein(GFP),red fluorescent protein(RFP)and improved light-oxygen-voltage(iLOV)genes were inserted into the restriction enzyme sites.Transfection of the constructs carrying the reporter genes into BHK-21 cells proved the rescue of the recombinant reporter viruses.Taken together,the establishment of a reverse genetic system for GETV provides a valuable tool for the study of the virus life cycle,and to aid the development of genetically engineered GETVs as vectors for foreign gene expression.展开更多
Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and l...Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation. We developed a cost-effective, rapid, and highly sensitive one-step "triplex RT-PCR enzyme hybridization" assay for simultaneous detections of Japanese Encephallitis virus (JEV, Flaviviridae), Getah virus (GETV, Togaviridae), and Tahyna virus (TAHV, Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction. The analytical sensitivity of this assay was 1 PFU/mL for JEV, 10 PFU/mL for GETV, and 10 PFU/mL for TAHV. This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods. When "triplex RT-PCR enzyme hybridization" was applied to 29 cerebrospinal fluid (CSF) samples that were JEV-positive by normal RT-PCR assay, all samples were strongly positive for JEV, but negative for GETV and TAHV, demonstrating a good sensitivity, specificity, and performance at CSF specimen detection.展开更多
基金supported by the National Program on Key Research Project of China(2018YFD0500104 and 2018YFD0500803)Technologies for Prevention and Control of Virus Zoonoses,Chinese Academy of Medical Sciences(2020-12M-5-001)。
文摘The Getah virus(GETV),a mosquito-borne RNA virus,is widely distributed in Oceania and Asia.GETV is not the only pathogenic to horses,pigs,cattle,foxes and boars,but it can also cause fever in humans.Since its first reported case in Chinese mainland in 2017,the number of GETV-affected provinces has increased to seventeen till now.Therefore,we performed an epidemiologic investigation of GETV in the Xinjiang region,located in northwestern China,during the period of 2017-2020.ELISA was used to analyze 3299 serum samples collected from thoroughbred horse,local horse,sheep,goat,cattle,and pigs,with thoroughbred horse(74.8%),local horse(67.3%),goat(11.7%),sheep(10.0%),cattle(25.1%)and pigs(51.1%)being positive for anti-GETV antibodies.Interestingly,the neutralizing antibody titer in horses was much higher than in other species.Four samples from horses and pigs were positive for GETV according to RT-PCR.Furthermore,from the serum of a local horse,we isolated GETV which was designated as strain XJ-2019-07,and determined its complete genome sequence.From the phylogenetic relationships,it belongs to the Group III lineage.This is the first evidence of GETV associated to domestic animals in Xinjiang.Overall,GETV is prevalent in Xinjiang and probably has been for several years.Since no vaccine against GETV is available in China,detection and monitoring strategies should be improved in horses and pigs,especially imported and farmed,in order to prevent economic losses.
基金funded by the Natural Science Foundation of Guangxi Province(No.2018GXNSFDA281021)the Foundation of Guangxi University(No.XGZ130959)。
文摘Getah virus(GETV)is a mosquito-borne virus of the genus Alphavirus in the family Togaviridae and,in recent years,it has caused several outbreaks in animals.The molecular basis for GETV pathogenicity is not well understood.Therefore,a reverse genetic system of GETV is needed to produce genetically modified viruses for the study of the viral replication and its pathogenic mechanism.Here,we generated a CMV-driven infectious cDNA clone based on a previously isolated GETV strain,GX201808(pGETV-GX).Transfection of pGETV-GX into BHK-21 cells resulted in the recovery of a recombinant virus(rGETV-GX)which showed similar growth characteristics to its parental virus.Then three-day-old mice were experimentally infected with either the parental or recombinant virus.The recombinant virus showed milder pathogenicity than the parental virus in the mice.Based on the established CMV-driven cDNA clone,subgenomic promoter and two restriction enzyme sites(BamHI and EcoRI)were introduced into the region between E1 protein and 3’UTR.Then the green fluorescent protein(GFP),red fluorescent protein(RFP)and improved light-oxygen-voltage(iLOV)genes were inserted into the restriction enzyme sites.Transfection of the constructs carrying the reporter genes into BHK-21 cells proved the rescue of the recombinant reporter viruses.Taken together,the establishment of a reverse genetic system for GETV provides a valuable tool for the study of the virus life cycle,and to aid the development of genetically engineered GETVs as vectors for foreign gene expression.
基金NIH Grant (2U54AI057160-06)Development Grant of State Key Laboratory for Infectious Disease Prevention and Control (2008SKLID105)
文摘Arboviruses represent a serious problem to public health and agriculture worldwide. Fast, accurate identification of the viral agents of arbovirus-associated disease is essential for epidemiological surveillance and laboratory investigation. We developed a cost-effective, rapid, and highly sensitive one-step "triplex RT-PCR enzyme hybridization" assay for simultaneous detections of Japanese Encephallitis virus (JEV, Flaviviridae), Getah virus (GETV, Togaviridae), and Tahyna virus (TAHV, Bunyaviridae) using three pairs of primers to amplify three target sequences in one RT-PCR reaction. The analytical sensitivity of this assay was 1 PFU/mL for JEV, 10 PFU/mL for GETV, and 10 PFU/mL for TAHV. This assay is significantly more rapid and less expensive than the traditional serological detection and single RT-PCR reaction methods. When "triplex RT-PCR enzyme hybridization" was applied to 29 cerebrospinal fluid (CSF) samples that were JEV-positive by normal RT-PCR assay, all samples were strongly positive for JEV, but negative for GETV and TAHV, demonstrating a good sensitivity, specificity, and performance at CSF specimen detection.
