Severe fever with thrombocytopenia syndrome(SFTS) is an emerging hemorrhagic fever in rural areas of China and is caused by a new bunyavirus,SFTSV,named after the disease.The transmission vectors and animal hosts of S...Severe fever with thrombocytopenia syndrome(SFTS) is an emerging hemorrhagic fever in rural areas of China and is caused by a new bunyavirus,SFTSV,named after the disease.The transmission vectors and animal hosts of SFTSV are unclear.Ticks are the most likely transmission vectors and domestic animals,including goats,dogs,and cattle,are potential amplifying hosts of SFTSV.The clinical symptoms of SFTS are nonspecific,but major symptoms include fever,gastrointestinal symptoms,myalgia,dizziness,joint pain,chills,and regional lymphadenopathy.The most common abnormalities in laboratory test results are thrombocytopenia(95%),leukocytopenia(86%),and elevated levels of serum alanine aminotransferase,aspartate aminotransferase,creatine kinase,and lactate dehydrogenase.The fatality rate for SFTS is 12% on average,and the annual incidence of the disease is approximately five per 100000 of the rural population.展开更多
Hubei Province is a major epidemic area of severe fever with thrombocytopenia syndrome bunyavirus(SFTSV) in China. However, to date, a few SFTSV strains have been isolated from Hubei Province, preventing effective stu...Hubei Province is a major epidemic area of severe fever with thrombocytopenia syndrome bunyavirus(SFTSV) in China. However, to date, a few SFTSV strains have been isolated from Hubei Province, preventing effective studies of epidemic outbreaks. Here, we report three confirmed patients(2015–2016) with typical symptoms of severe fever with thrombocytopenia syndrome disease(SFTS) who were farmers resident in different regions in Hubei Province. Three new SFTSV strains were isolated from the serum samples of each patient. Characterization of viral growth properties showed that there were no significant differences in virus production. All strains were completely sequenced, and phylogenetic analysis showed that unlike the other strains from Hubei province, which belonged to the SFTSV C3 genotype, one of the three strains belonged to the SFTSV C2 genotype. These results suggested that multiple SFTSV genotypes have been circulating in Hubei Province, providing insights into SFTSV evolution and improving our understanding of SFTSV prevalence in Hubei Province.展开更多
CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutio...CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science.Here,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant species.We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts.The resulting non-insect-transmissible viral vectors enabled effective and stable in planta delivery of Cas12a and Cas9 nucleases as well as adenine and cytosine base editors.In systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency.Plants with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic selection.Moreover,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable mutations.Because many plants are recalcitrant to stable transformation,the viral delivery system developed in this work provides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.展开更多
本文旨在探讨吉林省延边地区自然环境和动物体表蜱类分布及其消长规律,掌握其携带发热伴血小板减少综合征布尼亚病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)状况及传播病毒能力。2016年4~9月份按月采集延边州所辖8...本文旨在探讨吉林省延边地区自然环境和动物体表蜱类分布及其消长规律,掌握其携带发热伴血小板减少综合征布尼亚病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)状况及传播病毒能力。2016年4~9月份按月采集延边州所辖8个县(市)自然环境中生长的蜱虫和放牧动物体表蜱虫进行形态学分类,对其进行构成分析。对其中部分蜱虫分组进行SFTSV核酸检测。SFTSV核酸检测方法采用了Real time RT-PCR和RT-PCR相结合的方法。结果共采集蜱虫3 446只,其中森林革蜱763只,嗜群血蜱222只,日本血蜱639只,长角血蜱515只,全沟硬蜱1 014只,其他血蜱293只;全沟硬蜱(29.43%)和森林革蜱(22.14%)为本地优势种。长角血蜱在图们市(70.88%)和珲春市(40.59%)分布较多。对部分蜱虫(1605只)SFTSV Real time RT-PCR检测结果总最低感染率为1.81%,分种最低感染率分别为嗜群血蜱8.65%、日本血蜱4.53%、长角血蜱1.59%。序列分析结果表明,本文检测到的SFTSV病毒与我国其他省份从患者身上分离到的大部分SFTSV有高度一致性(99%)且与2012年从浙江患者血清中分离到的SFTSV、2013年韩国国家疾病预防控制中心从人身上采集的长角血蜱中分离到的SFTSV处于同一分枝,把该病毒命名为YBHC-TICK1-2016/CHINA。展开更多
Severe fever with thrombocytopenia syndrome virus （SFTSV） is a globe-shaped virus covered by a dense icosahedral array of glycoproteins Gn/Gc that mediate the attachment of the virus to host cells and the fusion of ...Severe fever with thrombocytopenia syndrome virus （SFTSV） is a globe-shaped virus covered by a dense icosahedral array of glycoproteins Gn/Gc that mediate the attachment of the virus to host cells and the fusion of viral and cellular membranes. Several membrane factors are involved in virus entry, including C-type lectins and nonmuscle myosin heavy chain IIA. The post-fusion crystal structure of the Gc protein suggests that it is a class II membrane fusion protein, similar to the E/E1 protein of flaviviruses and alphaviruses. The virus particles are internalized into host cell endosomes through the clathrin-dependent pathway, where the low pH activates the fusion of the virus with the cell membrane. With information from studies on other bunyaviruses, herein we will review our knowledge of the entry process of SFTSV.展开更多
目的分析济南市2011-2013年发热伴血小板减少综合征(severe fever with thrombocytopenia syndrome,SFTS)的流行病学特征和病原学特性,为更好地预防和控制SFTS提供科学依据。方法采用实时荧光定量RT-PCR方法检测济南市2011-2013年SFTS...目的分析济南市2011-2013年发热伴血小板减少综合征(severe fever with thrombocytopenia syndrome,SFTS)的流行病学特征和病原学特性,为更好地预防和控制SFTS提供科学依据。方法采用实时荧光定量RT-PCR方法检测济南市2011-2013年SFTS疑似病例急性期血清样本和蜱虫样本中的SFTS布尼亚病毒核酸,采用RT-PCR方法进行扩增并测序,使用DNASTAR和MEGA软件对测序结果进行序列比对和进化分析;用描述性流行病学方法对监测数据进行统计分析。结果 2011-2013年济南市共确诊95例SFTS病例,检测出3只蜱虫样本中的SFTS布尼亚病毒核酸阳性;发病地区主要集中在章丘市。对SFTS布尼亚病毒的S片段基因进行测序,结果显示2012年SFTS布尼亚病毒基因与安徽、江苏、河南分离株遗传距离相对较近。2013年SFTS布尼亚病毒基因与山东本地分离株遗传距离相对较近。结论济南市发热伴血小板减少综合征病例以新布尼亚病毒为其主要病原;夏秋季多发;中老年农民为主,具有明显职业性;发病范围逐年扩大,局部地区疫情活跃。应加强传染源的管理控制,深入开展对高危人群宣传教育和行为干预。展开更多
Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology ...Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology involved in SFTSV infection remains obscure.There are seven major genotypes of SFTSV(C1-C4 and J1-J3)and previously a reverse genetic system was established on a C3 strain of SFTSV.Here,we reported successfully establishment of a reverse genetics system based on a SFTSV C4 strain.First,we obtained the 5’-and 3’-terminal untranslated region(UTR)sequences of the Large(L),Medium(M)and Small(S)segments of a laboratory-adapted SFTSV C4 strain through rapid amplification of cDNA ends analysis,and developed functional T7 polymerase-based L-,M-and S-segment minigenome assays.Then,fulllength cDNA clones were constructed and infectious SFTSV were recovered from co-transfected cells.Viral infectivity,growth kinetics,and viral protein expression profile of the rescued virus were compared with the laboratory-adapted virus.Focus formation assay showed that the size and morphology of the foci formed by the rescued SFTSV were indistinguishable with the laboratory-adapted virus.However,one-step growth curve and nucleoprotein expression analyses revealed the rescued virus replicated less efficiently than the laboratory-adapted virus.Sequence analysis indicated that the difference may be due to the mutations in the laboratory-adapted strain which are more prone to cell culture.The results help us to understand the molecular biology of SFTSV,and provide a useful tool for developing vaccines and antivirals against SFTS.展开更多
文摘Severe fever with thrombocytopenia syndrome(SFTS) is an emerging hemorrhagic fever in rural areas of China and is caused by a new bunyavirus,SFTSV,named after the disease.The transmission vectors and animal hosts of SFTSV are unclear.Ticks are the most likely transmission vectors and domestic animals,including goats,dogs,and cattle,are potential amplifying hosts of SFTSV.The clinical symptoms of SFTS are nonspecific,but major symptoms include fever,gastrointestinal symptoms,myalgia,dizziness,joint pain,chills,and regional lymphadenopathy.The most common abnormalities in laboratory test results are thrombocytopenia(95%),leukocytopenia(86%),and elevated levels of serum alanine aminotransferase,aspartate aminotransferase,creatine kinase,and lactate dehydrogenase.The fatality rate for SFTS is 12% on average,and the annual incidence of the disease is approximately five per 100000 of the rural population.
基金supported by the Science and Technology Basic Work Program (2013FY113500)the National Key Research and Development Program (2016YFE 0113500) from the Ministry of Science and Technology of Chinathe European Union’s Horizon 2020 EVAg project (No 653316)
文摘Hubei Province is a major epidemic area of severe fever with thrombocytopenia syndrome bunyavirus(SFTSV) in China. However, to date, a few SFTSV strains have been isolated from Hubei Province, preventing effective studies of epidemic outbreaks. Here, we report three confirmed patients(2015–2016) with typical symptoms of severe fever with thrombocytopenia syndrome disease(SFTS) who were farmers resident in different regions in Hubei Province. Three new SFTSV strains were isolated from the serum samples of each patient. Characterization of viral growth properties showed that there were no significant differences in virus production. All strains were completely sequenced, and phylogenetic analysis showed that unlike the other strains from Hubei province, which belonged to the SFTSV C3 genotype, one of the three strains belonged to the SFTSV C2 genotype. These results suggested that multiple SFTSV genotypes have been circulating in Hubei Province, providing insights into SFTSV evolution and improving our understanding of SFTSV prevalence in Hubei Province.
基金supported by grants from the National Key R&D Program of China(2022YFC2601000)the Natural Science Foundation of Zhejiang Province,China(no.LZ20C140004)the National Natural Science Foundation of China(no.31870142).
文摘CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science.Here,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant species.We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts.The resulting non-insect-transmissible viral vectors enabled effective and stable in planta delivery of Cas12a and Cas9 nucleases as well as adenine and cytosine base editors.In systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency.Plants with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic selection.Moreover,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable mutations.Because many plants are recalcitrant to stable transformation,the viral delivery system developed in this work provides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.
文摘本文旨在探讨吉林省延边地区自然环境和动物体表蜱类分布及其消长规律,掌握其携带发热伴血小板减少综合征布尼亚病毒(Severe fever with thrombocytopenia syndrome virus,SFTSV)状况及传播病毒能力。2016年4~9月份按月采集延边州所辖8个县(市)自然环境中生长的蜱虫和放牧动物体表蜱虫进行形态学分类,对其进行构成分析。对其中部分蜱虫分组进行SFTSV核酸检测。SFTSV核酸检测方法采用了Real time RT-PCR和RT-PCR相结合的方法。结果共采集蜱虫3 446只,其中森林革蜱763只,嗜群血蜱222只,日本血蜱639只,长角血蜱515只,全沟硬蜱1 014只,其他血蜱293只;全沟硬蜱(29.43%)和森林革蜱(22.14%)为本地优势种。长角血蜱在图们市(70.88%)和珲春市(40.59%)分布较多。对部分蜱虫(1605只)SFTSV Real time RT-PCR检测结果总最低感染率为1.81%,分种最低感染率分别为嗜群血蜱8.65%、日本血蜱4.53%、长角血蜱1.59%。序列分析结果表明,本文检测到的SFTSV病毒与我国其他省份从患者身上分离到的大部分SFTSV有高度一致性(99%)且与2012年从浙江患者血清中分离到的SFTSV、2013年韩国国家疾病预防控制中心从人身上采集的长角血蜱中分离到的SFTSV处于同一分枝,把该病毒命名为YBHC-TICK1-2016/CHINA。
基金funded by the National Key Plan for Scientific Research and Development of China (2016YFD0500300)the Strategic Priority Research Program of the Chinese Academy of Sciences (CAS) (XDB11030800)the Natural Science Foundation of China (L1524009)
文摘Severe fever with thrombocytopenia syndrome virus （SFTSV） is a globe-shaped virus covered by a dense icosahedral array of glycoproteins Gn/Gc that mediate the attachment of the virus to host cells and the fusion of viral and cellular membranes. Several membrane factors are involved in virus entry, including C-type lectins and nonmuscle myosin heavy chain IIA. The post-fusion crystal structure of the Gc protein suggests that it is a class II membrane fusion protein, similar to the E/E1 protein of flaviviruses and alphaviruses. The virus particles are internalized into host cell endosomes through the clathrin-dependent pathway, where the low pH activates the fusion of the virus with the cell membrane. With information from studies on other bunyaviruses, herein we will review our knowledge of the entry process of SFTSV.
基金supported by grants from the National Natural Science Foundation of China(No.31900146Open Research Fund Program of the State Key Laboratory of Virology of China(No.2020IOV003)Team project of Health Commission of Hubei Province(WJ2019C003)。
文摘Severe fever with thrombocytopenia syndrome virus(SFTSV)is an emerging tick-borne bunyavirus that causes hemorrhagic fever-like disease(SFTS)in humans with a case fatality rate up to 30%.To date,the molecular biology involved in SFTSV infection remains obscure.There are seven major genotypes of SFTSV(C1-C4 and J1-J3)and previously a reverse genetic system was established on a C3 strain of SFTSV.Here,we reported successfully establishment of a reverse genetics system based on a SFTSV C4 strain.First,we obtained the 5’-and 3’-terminal untranslated region(UTR)sequences of the Large(L),Medium(M)and Small(S)segments of a laboratory-adapted SFTSV C4 strain through rapid amplification of cDNA ends analysis,and developed functional T7 polymerase-based L-,M-and S-segment minigenome assays.Then,fulllength cDNA clones were constructed and infectious SFTSV were recovered from co-transfected cells.Viral infectivity,growth kinetics,and viral protein expression profile of the rescued virus were compared with the laboratory-adapted virus.Focus formation assay showed that the size and morphology of the foci formed by the rescued SFTSV were indistinguishable with the laboratory-adapted virus.However,one-step growth curve and nucleoprotein expression analyses revealed the rescued virus replicated less efficiently than the laboratory-adapted virus.Sequence analysis indicated that the difference may be due to the mutations in the laboratory-adapted strain which are more prone to cell culture.The results help us to understand the molecular biology of SFTSV,and provide a useful tool for developing vaccines and antivirals against SFTS.
