Monkeypox virus(MPXV),a DNA virus belonging to the Orthopoxvirus genus,causes a self-limiting zoonotic disease known as mpox.The human monkeypox infection was first recorded in a 9-month-old child in the Republic of C...Monkeypox virus(MPXV),a DNA virus belonging to the Orthopoxvirus genus,causes a self-limiting zoonotic disease known as mpox.The human monkeypox infection was first recorded in a 9-month-old child in the Republic of Congo in the 1970s(Ladnyj et al.,1972).For a long time,mpox was mainly prevalent in the humid forests of Central Africa and parts of West Africa(Kabugae and El Zowalaty,2019).In recent years,MPXV has been spread to other countries through trade and travel.In 2003,a group of rare pets carrying MPXV was exported from Africa to the United States,and an outbreak of animal-to-human transmission occurred(Di Giulio and Eckburg,2004).In 2018,two cases of mpox were confirmed in travelers from Nigeria to the United Kingdom and one case was confirmed in Israel(Vaughan et al.,2018;Erez et al.,2019).Then,in 2019,one case was confirmed in Singapore(Ng et al.,2019).The outbreak of mpox in multiple non-endemic countries in North America and Europe started in May 2022(WHO,2022b).On July 23,2022,the WHO declared MPXV a Public Health Emergency of International Concern(PHEIC)(WHO,2022a).As of March 5,2023,86,309 confirmed mpox cases have been reported from 107 countries/regions worldwide(WHO,2023).In the mainland of China,the first imported case was confirmed by the Chinese Center for Disease Control and Prevention(Zhao et al.,2022),making this the fifth confirmed mpox infection in China.Neglected zoonotic mpox has been restricted in Africa but now it is back in the spotlight worldwide(Tan and Gao,2022).展开更多
The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary...The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads,as it is one of the DNA viruses with the largest genome and the most AT-biased,and has a significant number of tandem repeats.Here we evaluated the performance of metagenomic and amplicon sequencing techniques,and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland.We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens.Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes.Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences.Besides,several intra-host single nucleotide variations were identified in the first imported mpox case.This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens.The findings of this study will significantly enhance the surveillance of MPXV.展开更多
Since May 2022,a severe global Mpox epidemic has underscored the urgent need for a preventative vaccine.On September 16,2022,the mainland of China reported its first case of imported Mpox,which was subsequently follow...Since May 2022,a severe global Mpox epidemic has underscored the urgent need for a preventative vaccine.On September 16,2022,the mainland of China reported its first case of imported Mpox,which was subsequently followed by a significant rise in domestic infections commencing from June 2023.This alarming trend has escalated the likelihood of localized outbreaks and covert transmission,posing a heightened risk to public health.Notably,the United States,many European countries,and Japan have approved the use of smallpox vaccines for Mpox prevention and emergency vaccination post-exposure,based on their cross-protection efficacy.In recent years,virology research has broadened its scope to include investigations into various novel vaccine approaches,such as nucleic acid-based vaccines,protein subunit vaccines,and epitope peptide vaccines,and other related methodologies.This review offers a thorough examination of the current global landscape of Mpox prevalence,delves into the advancements in Mpox vaccine development,and highlights the progress achieved in Mpox vaccine research,serving as a valuable resource and providing technical insights essential for the effective prevention and control of Mpox.展开更多
Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investi...Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant(lineage B.1.351).Methods K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant.Viral replication,pathological phenotypes,and brain transcriptomes were analyzed.Gene Ontology(GO)analysis was performed to identify altered pathways.Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot.Results Pathological alterations were observed in the lungs of both mouse strains.However,only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection,accompanied by neuropathological injury and weight loss.GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses,including type I interferons,pro-inflammatory cytokines,Toll-like receptor signaling components,and interferon-stimulated genes.Neuroinflammation was evident,alongside activation of apoptotic and pyroptotic pathways.Furthermore,altered neural cell marker expression suggested viral-induced neuroglial activation,resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks.Conclusion These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.展开更多
Background:Vaccinia virus(VACV)and mpox virus(MPXV)belong to the orthopoxvirus genus and share high genetic similarity,making VACV widely used in the mpox pandemic.CAST/EiJ mice have been widely used for studying orth...Background:Vaccinia virus(VACV)and mpox virus(MPXV)belong to the orthopoxvirus genus and share high genetic similarity,making VACV widely used in the mpox pandemic.CAST/EiJ mice have been widely used for studying orthopoxvirus infection.However,the histopathological features of CAST/EiJ mice with mpox virus(MPXV)and vaccinia virus(VACV)infections have not been fully elucidated.Methods:Four group of CAST/EiJ mice were challenged with low-dose VACV(103 PFU,VACV-L),high-dose VACV(106 PFU,VACV-H),MPXV(106 PFU)or PBS via intraperitoneal route,and the disease signs and body weight were monitored daily.Subsequently,viral loads and titers in the blood and spleen of CAST/EiJ mice were analyzed via qPCR and TCID 50 assay.Finally,the spleen samples were analyzed for histopathological,immunohistochemical and RNA-seq.Results:Herein,we found that VACV-L and MPXV caused splenomegaly via the intraperitoneal route,whereas VACV-H caused rapid lethality with limited splenomegaly.Transcriptome analysis from spleen revealed significant differences in gene expression between VACV-L and VACV-H groups,but the differentially expressed genes induced by splenomegaly between VACV-L and MPXV groups were highly similar.Furthermore,pathway enrichment analysis demonstrated that the VACV-L,VACV-H,and MPXV groups were all associated with the calcium,MAPK,and PI3K-Akt signaling pathway.Compared to the lethal infection observed in VACV-H group,the splenomegaly in the VACV-L and MPXV groups was characterized by extramedullary hematopoiesis and increased macrophages infiltration in the red pulp.Transcriptome analysis of the spleen demonstrated that the Wnt,tumor necrosis factor(TNF),and transforming growth factorβ(TGF-β)signaling pathways may promote splenomegaly by modulating granulocyte infiltration and inflammatory responses.Compared to VACV-L group,the limited splenomegaly but lethality in VACV-H-infected mice might be associated with extensive splenic necrosis,diffuse congestion,and hemorrhage in the red pulp,as well as changes in the cGMP-PKG,Ras signaling,and Fc gamma Rmediated phagocytosis pathways.Conclusions:Our findings systematically compared the pathogenicity of VACV and MPXV in CAST/EiJ mice,incorporating splenic transcriptome analysis to provide insights into the potential molecular mechanism behind orthopoxvirus-induced splenomegaly in CAST/EiJ mice.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
The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November ...The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November 20,2020,51 detection kits for SARS-CoV-2 nucleic acids(24 kits),antibodies(25 kits),or antigens(2 kits)have been approved by the National Medical Products Administration of China(NMPA).Convenient and reliable SARS-CoV-2 detection assays are urgently needed worldwide for strategic control of the pandemic.In this review,the detection kits approved in China are summarised and the three types of tests,namely nucleic acid,serological and antigen detection,which are available for the detection of COVID-19 are discussed in detail.The development of novel detection kits will lay the foundation for the control and prevention of the COVID-19 pandemic globally.展开更多
Objective Tick-borne encephalitis virus(TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis(TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease....Objective Tick-borne encephalitis virus(TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis(TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. Methods A reverse-transcription recombinase-aided amplification(RT-RAA) assay was developed. This assay can be completed in one closed tube at 39℃ within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type(WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. Results The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units(pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. Conclusion A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.展开更多
Pseudorabies virus(PRV),a veterinary pathogen that infects domestic animals as well as wild animals such as wild boar and feral swine,was recently reported to infect human and led to endophthalmitis and encephalitis.A...Pseudorabies virus(PRV),a veterinary pathogen that infects domestic animals as well as wild animals such as wild boar and feral swine,was recently reported to infect human and led to endophthalmitis and encephalitis.A retrospective seroepidemiologic survey was conducted using 1,335 serum samples collected from patients with encephalitis and ELISA positive rates were 12.16%,14.25%,and 6.52%in 2012,2013,and 2017,respectively.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
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
Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infection...Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infections. Coxsackievirus B6 (CV-B6)belongs to the species EV-B, which currently consists of 63 serotypes, including echovirus (serotypes 1–7,9, 11–21, 24–27, 29–33), coxsackievirus group A (CVA9), coxsackievirus group B (CV-B, serotypes 1–6),the newly identified EVs (serotypes EV-B69, B73–75,B77–88, B93, B97–98, B100–101, B106–107, and B110–113), and the simian enterovirus SA5.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
Zika virus(ZIKV) causes rash, moderate fever, conjunctivitis, and arthralgia, and has serious connection with neurological complications;therefore, it is a major threat to public health. A rapid and supersensitive met...Zika virus(ZIKV) causes rash, moderate fever, conjunctivitis, and arthralgia, and has serious connection with neurological complications;therefore, it is a major threat to public health. A rapid and supersensitive method for detecting anti-ZIKV antibodies in humans and animals is thus urgently required. Here, we report an NS1-based luciferase immunosorbent assay(LISA), developed to detect ZIKV-specific IgG. Fusion proteins including a reporter Nano-luciferase(NLuc) and various fragments of ZIKV NS1 protein were expressed in 293 T cells. LISA was performed using the above cell lysates containing the expressed fusion proteins. Sample panels of humans and animals infected with ZIKV were examined for sensitivity of LISA, relative to those of ZIKV RT-PCR, commercial NS1-based ELISA, and micro-neutralization(MN) assays.Specificity and potential cross-reactivity were also evaluated using various convalescent serum samples derived from patients infected with dengue virus(DENV), Japanese encephalitis virus(JEV), and hepatitis C virus(HCV). Results indicated the optimal antigenic domain for anti-ZIKV IgG detection was located within 172–352 amino acids(aa) of ZIKV NS1 protein. NS1-based LISA performs better than commercial ELISA in anti-ZIKV Ig G detection. LISA was shown to be at least fourfold more sensitive than commercial ELISA, and could detect anti-ZIKV Ig G in various animal hosts without the need of species-specific labeled antibody. This novel assay is potentially useful for the rapid and sensitive detection of anti-ZIKV IgG in human and animal samples.展开更多
Objective The current outbreak of Zika virus(ZIKV)poses a severe threat to human health.Two ZIKV strains were isolated from mosquitoes collected from the Dejiang prefecture in China in 2016,which was the first isolati...Objective The current outbreak of Zika virus(ZIKV)poses a severe threat to human health.Two ZIKV strains were isolated from mosquitoes collected from the Dejiang prefecture in China in 2016,which was the first isolation of ZIKV in nature in China.Methods In this study,serum samples were collected from 366 healthy individuals and 104 animals from Dejiang prefecture in 2017,and the plaque reduction neutralization test(PRNT)was used to evaluate the seroprevalence of ZIKV.Results None of the 366 residents from whom the samples were collected were seropositive for ZIKV.None of the 11 pigs from whom the samples were collected were seropositive for ZIKV,while 1 of 63(1.59%)chickens and 2 of 30(6.67%)sheep were seropositive for ZIKV.Conclusions The extremely low seropositivity rate of ZIKV antibodies in animals in the Dejiang prefecture,Guizhou province in this study indicates that ZIKV can infect animals;however,there is a low risk of ZIKV circulating in the local population.展开更多
West Nile virus(WNV)is an important neurotropic flavivirus that is widely distributed globally.WNV strain XJ11129 was first isolated in Xinjiang,China,and its genetic and biological characteristics remain largely unkn...West Nile virus(WNV)is an important neurotropic flavivirus that is widely distributed globally.WNV strain XJ11129 was first isolated in Xinjiang,China,and its genetic and biological characteristics remain largely unknown.In this study,phylogenetic and sequence analyses revealed that XJ11129 belongs to lineage 1 a and shares high genetic identity with the highly pathogenic strain NY99.Then,the full-length genomic c DNA of XJ11129 was amplified and assembled using a modified Gibson assembly(GA)method.The virus(named r XJ11129)was successfully rescued in days following this method.Compared with other wild-type WNV isolates,r XJ11129 exhibited virulence indistinguishable from that of the NY99 strain in vivo.In summary,the genomic and virulence phenotypes of r XJ11129 were characterized in vivo and in vitro,and these data will improve the understanding of the spread and pathogenesis of this reemerging virus.展开更多
Dear Editor,A novel coronavirus disease(COVID-19)caused by acute respiratory syndrome coronavirus 2(SARS-Co V-2)first broke out in Wuhan,Hubei Province,China,in 2019.More than 50 million cases of COVID-19 have been co...Dear Editor,A novel coronavirus disease(COVID-19)caused by acute respiratory syndrome coronavirus 2(SARS-Co V-2)first broke out in Wuhan,Hubei Province,China,in 2019.More than 50 million cases of COVID-19 have been confirmed globally with 1,280,868 deaths reported as of November12,2020(https://covid19.who.int/).展开更多
Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMR...Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMRV-GCCDC7),in the Qinghai-Tibet Plateau wild pika(Ochotona curzoniae).Though the PMRVGCCDC7 shows features of a typical reovirus with ten gene segments arranged in 3:3:4 in length,the virus belongs to an independent evolutionary branch compared to other MRVs based on phylogenetic tree analysis.The results of cellular susceptibility,species tropism,and replication kinetics of PMRV-GCCDC7 indicated the virus could infect four human cell lines(A549,Huh7,HCT,and LoVo)and six non-human cell lines,including Vero-E6,LLCMK2,BHK-21,N2a,MDCK,and RfKT cell,derived from diverse mammals,i.e.monkey,mice,canine and bat,which revealed the potential of PMRV-GCCDC7 to infect a variety of hosts.Infection of BALB/c mice with PMRVGCCDC7 via intranasal inoculation led to relative weight loss,lung tissue damage and inflammation with the increase of virus titer,but no serious respiratory symptoms and death occurred.The characterization of the new reovirus from a plateau-based wild animal has expanded our knowledge of the host range of MRV and provided insight into its risk of trans-species transmission and zoonotic diseases.展开更多
Background:This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1.Methods:Humanized fragments,consisting of the endothelial cell-specific K18 promoter,human ST6GAL1-encodi...Background:This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1.Methods:Humanized fragments,consisting of the endothelial cell-specific K18 promoter,human ST6GAL1-encoding gene,and luciferase gene,were microinjected into the fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offspring were identified using PCR.Mice exhibiting elevated expression of the hST6GAL1 gene were selectively bred for propagation,and in vivo analysis was performed for screening.Expression of the humanized gene was tested by performing immunohistochemical(IHC)analysis.Hematologic and biochemical analyses using the whole blood and serum of humanized hST6GAL1 mice were performed.Results:Successful integration of the human ST6GAL1 gene into the mouse genome led to the overexpression of human SiaT ST6GAL1.Seven mice were identified as carrying copies of the humanized gene,and the in vivo analysis indicated that hST6GAL1gene expression in positive mice mirrored influenza virus infection characteristics.The IHC results revealed that hST6GAL1 was expressed in the lungs of humanized mice.Moreover,the hematologic and biochemical parameters of the positive mice were within the normal range.Conclusion:A humanized influenza mouse model expressing the hST6GAL1 gene was successfully established and characterized.展开更多
TheRhabdoviridae family comprises a diverse range of negative-sense single-stranded ribonucleic acid(RNA)viruses,including significant human and mammalian viruses transmitted by various arthropod species.Herein,usingA...TheRhabdoviridae family comprises a diverse range of negative-sense single-stranded ribonucleic acid(RNA)viruses,including significant human and mammalian viruses transmitted by various arthropod species.Herein,usingAedes albopictus(Ae.albopictus)samples collected in two urban parks during 2023 and 2024,through metagenomics sequencing,16 sequences were identified as putative novel viruses,showing closest homology to insect-specific viruses,mycoviruses,or plant-associated viruses.Notably,two novel viruses,Aedes albopictus almendravirus GCCDC15(Aealb-AlmV GCCDC15)and Aedes albopictus almendravirus GCCDC16(Aealb-AlmV GCCDC16)were identified and successfully isolated.Both of these viruses belong to the genusAlmendravirus within theRhabdoviridae family.Phylogenetic analysis revealed that Aealb-AlmV GCCDC15 and GCCDC16 are distantly related to Coot Bay virus(the United States of America,2013)and Menghai rhabdovirus(Yunnan Province,China,2017).The genetic distances between these two viruses and their most similar viruses are marked by 59.85%and 87.20%of amino acid identity in the L protein,respectively,supporting their classification as two new species in theRhabdoviridae family.Cytopathic effects and rod-like virions were observed in mosquito cells(C6/36)after inoculating with supernatants from theAe.albopictus samples.To investigate the natural distribution and persistence of the novel almendraviruses,we conducted a specific reverse transcription-polymerase chain reaction(RT-PCR)screening ofAe.albopictus mosquitoes collected from two urban parks across different time points.The assays confirmed the presence of both Aealb-AlmV GCCDC15 and GCCDC16 in mosquito populations.Critically,these viruses were detected repeatedly over successive sampling periods and in mosquitoes from geographically distinct sites within the urban environment.In summary,our study delineates the virome characteristics ofAedes mosquitoes in the urban ecosystem and successfully isolated two novel rhabdoviruses.The recurrent detection provides clear evidence for the sustained circulation ofAe.albopictus-derived almendraviruses in urban parks,highlighting their ongoing transmission and establishment in these habitats.展开更多
基金supported by funding from the National Key R&D Program of China"Risk Identification of Potential New Pathogens and Development of Broad-spectrum Antibodies"(2022YFC2303401).
文摘Monkeypox virus(MPXV),a DNA virus belonging to the Orthopoxvirus genus,causes a self-limiting zoonotic disease known as mpox.The human monkeypox infection was first recorded in a 9-month-old child in the Republic of Congo in the 1970s(Ladnyj et al.,1972).For a long time,mpox was mainly prevalent in the humid forests of Central Africa and parts of West Africa(Kabugae and El Zowalaty,2019).In recent years,MPXV has been spread to other countries through trade and travel.In 2003,a group of rare pets carrying MPXV was exported from Africa to the United States,and an outbreak of animal-to-human transmission occurred(Di Giulio and Eckburg,2004).In 2018,two cases of mpox were confirmed in travelers from Nigeria to the United Kingdom and one case was confirmed in Israel(Vaughan et al.,2018;Erez et al.,2019).Then,in 2019,one case was confirmed in Singapore(Ng et al.,2019).The outbreak of mpox in multiple non-endemic countries in North America and Europe started in May 2022(WHO,2022b).On July 23,2022,the WHO declared MPXV a Public Health Emergency of International Concern(PHEIC)(WHO,2022a).As of March 5,2023,86,309 confirmed mpox cases have been reported from 107 countries/regions worldwide(WHO,2023).In the mainland of China,the first imported case was confirmed by the Chinese Center for Disease Control and Prevention(Zhao et al.,2022),making this the fifth confirmed mpox infection in China.Neglected zoonotic mpox has been restricted in Africa but now it is back in the spotlight worldwide(Tan and Gao,2022).
基金supported by the National Key Research and Development Program of China(2022YFC2303401,2022YFC2304100,2016YFD0500301,2021YFC0863300)the Beijing Science and Technology Plan(Z211100002521017)the National Natural Science Foundation of China(82241080)。
文摘The monkeypox virus(MPXV)has triggered a current outbreak globally.Genome sequencing of MPXV and rapid tracing of genetic variants will benefit disease diagnosis and control.It is a significant challenge but necessary to optimize the strategy and application of rapid full-length genome identification and to track variations of MPXV in clinical specimens with low viral loads,as it is one of the DNA viruses with the largest genome and the most AT-biased,and has a significant number of tandem repeats.Here we evaluated the performance of metagenomic and amplicon sequencing techniques,and three sequencing platforms in MPXV genome sequencing based on multiple clinical specimens of five mpox cases in Chinese mainland.We rapidly identified the full-length genome of MPXV with the assembly of accurate tandem repeats in multiple clinical specimens.Amplicon sequencing enables cost-effective and rapid sequencing of clinical specimens to obtain high-quality MPXV genomes.Third-generation sequencing facilitates the assembly of the terminal tandem repeat regions in the monkeypox virus genome and corrects a common misassembly in published sequences.Besides,several intra-host single nucleotide variations were identified in the first imported mpox case.This study offers an evaluation of various strategies aimed at identifying the complete genome of MPXV in clinical specimens.The findings of this study will significantly enhance the surveillance of MPXV.
文摘Since May 2022,a severe global Mpox epidemic has underscored the urgent need for a preventative vaccine.On September 16,2022,the mainland of China reported its first case of imported Mpox,which was subsequently followed by a significant rise in domestic infections commencing from June 2023.This alarming trend has escalated the likelihood of localized outbreaks and covert transmission,posing a heightened risk to public health.Notably,the United States,many European countries,and Japan have approved the use of smallpox vaccines for Mpox prevention and emergency vaccination post-exposure,based on their cross-protection efficacy.In recent years,virology research has broadened its scope to include investigations into various novel vaccine approaches,such as nucleic acid-based vaccines,protein subunit vaccines,and epitope peptide vaccines,and other related methodologies.This review offers a thorough examination of the current global landscape of Mpox prevalence,delves into the advancements in Mpox vaccine development,and highlights the progress achieved in Mpox vaccine research,serving as a valuable resource and providing technical insights essential for the effective prevention and control of Mpox.
基金supported by the National Key Research and Development Program of China(2023YFC3041500).
文摘Objective Patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection frequently develop central nervous system damage,yet the mechanisms driving this pathology remain unclear.This study investigated the primary pathways and key factors underlying brain tissue damage induced by the SARS-CoV-2 beta variant(lineage B.1.351).Methods K18-hACE2 and C57BL/6 mice were intranasally infected with the SARS-CoV-2 beta variant.Viral replication,pathological phenotypes,and brain transcriptomes were analyzed.Gene Ontology(GO)analysis was performed to identify altered pathways.Expression changes of host genes were verified using reverse transcription-quantitative polymerase chain reaction and Western blot.Results Pathological alterations were observed in the lungs of both mouse strains.However,only K18-hACE2 mice exhibited elevated viral RNA loads and infectious titers in the brain at 3 days post-infection,accompanied by neuropathological injury and weight loss.GO analysis of infected K18-hACE2 brain tissue revealed significant dysregulation of genes associated with innate immunity and antiviral defense responses,including type I interferons,pro-inflammatory cytokines,Toll-like receptor signaling components,and interferon-stimulated genes.Neuroinflammation was evident,alongside activation of apoptotic and pyroptotic pathways.Furthermore,altered neural cell marker expression suggested viral-induced neuroglial activation,resulting in caspase 4 and lipocalin 2 release and disruption of neuronal molecular networks.Conclusion These findings elucidate mechanisms of neuropathogenicity associated with the SARS-CoV-2 beta variant and highlight therapeutic targets to mitigate COVID-19-related neurological dysfunction.
基金Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences,Grant/Award Number:2023-I2M-2-001National Key Research and Development Project of China,Grant/Award Number:2022YFC2304100 and 2023YFC2309000+1 种基金National Natural Science Foundation of China,Grant/Award Number:82241068 and 82222041Beijing Natural Science Foundation,Grant/Award Number:Z220018。
文摘Background:Vaccinia virus(VACV)and mpox virus(MPXV)belong to the orthopoxvirus genus and share high genetic similarity,making VACV widely used in the mpox pandemic.CAST/EiJ mice have been widely used for studying orthopoxvirus infection.However,the histopathological features of CAST/EiJ mice with mpox virus(MPXV)and vaccinia virus(VACV)infections have not been fully elucidated.Methods:Four group of CAST/EiJ mice were challenged with low-dose VACV(103 PFU,VACV-L),high-dose VACV(106 PFU,VACV-H),MPXV(106 PFU)or PBS via intraperitoneal route,and the disease signs and body weight were monitored daily.Subsequently,viral loads and titers in the blood and spleen of CAST/EiJ mice were analyzed via qPCR and TCID 50 assay.Finally,the spleen samples were analyzed for histopathological,immunohistochemical and RNA-seq.Results:Herein,we found that VACV-L and MPXV caused splenomegaly via the intraperitoneal route,whereas VACV-H caused rapid lethality with limited splenomegaly.Transcriptome analysis from spleen revealed significant differences in gene expression between VACV-L and VACV-H groups,but the differentially expressed genes induced by splenomegaly between VACV-L and MPXV groups were highly similar.Furthermore,pathway enrichment analysis demonstrated that the VACV-L,VACV-H,and MPXV groups were all associated with the calcium,MAPK,and PI3K-Akt signaling pathway.Compared to the lethal infection observed in VACV-H group,the splenomegaly in the VACV-L and MPXV groups was characterized by extramedullary hematopoiesis and increased macrophages infiltration in the red pulp.Transcriptome analysis of the spleen demonstrated that the Wnt,tumor necrosis factor(TNF),and transforming growth factorβ(TGF-β)signaling pathways may promote splenomegaly by modulating granulocyte infiltration and inflammatory responses.Compared to VACV-L group,the limited splenomegaly but lethality in VACV-H-infected mice might be associated with extensive splenic necrosis,diffuse congestion,and hemorrhage in the red pulp,as well as changes in the cGMP-PKG,Ras signaling,and Fc gamma Rmediated phagocytosis pathways.Conclusions:Our findings systematically compared the pathogenicity of VACV and MPXV in CAST/EiJ mice,incorporating splenic transcriptome analysis to provide insights into the potential molecular mechanism behind orthopoxvirus-induced splenomegaly in CAST/EiJ mice.
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 the National Key Program for Infectious Disease of China(2018ZX10101002)the National Key Research and Development Program of China(2016YFD0500301,2020YFC0840900)。
文摘The on-going global pandemic of coronavirus disease 2019(COVID-19)caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has been underway for about 11 months.Through November 20,2020,51 detection kits for SARS-CoV-2 nucleic acids(24 kits),antibodies(25 kits),or antigens(2 kits)have been approved by the National Medical Products Administration of China(NMPA).Convenient and reliable SARS-CoV-2 detection assays are urgently needed worldwide for strategic control of the pandemic.In this review,the detection kits approved in China are summarised and the three types of tests,namely nucleic acid,serological and antigen detection,which are available for the detection of COVID-19 are discussed in detail.The development of novel detection kits will lay the foundation for the control and prevention of the COVID-19 pandemic globally.
基金supported by the National key research and development project [2017YFC1200505]the National Science and Technology Major Project of China [2018ZX10711001,2018ZX10101-002]the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control [2015SKLID505,2014SKLID103]
文摘Objective Tick-borne encephalitis virus(TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis(TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. Methods A reverse-transcription recombinase-aided amplification(RT-RAA) assay was developed. This assay can be completed in one closed tube at 39℃ within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type(WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. Results The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units(pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. Conclusion A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype.
基金supported by Ten-thousand Talents Program [Dr.Xiangdong Li]National Key Research and Development Program 2018ZX10101002,2016YFD0500401+1 种基金the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control 2015SKLID505Scientific Research Project of China CDC JY18-1-01。
文摘Pseudorabies virus(PRV),a veterinary pathogen that infects domestic animals as well as wild animals such as wild boar and feral swine,was recently reported to infect human and led to endophthalmitis and encephalitis.A retrospective seroepidemiologic survey was conducted using 1,335 serum samples collected from patients with encephalitis and ELISA positive rates were 12.16%,14.25%,and 6.52%in 2012,2013,and 2017,respectively.
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 Key Science and Technology Projects of China [Project Nos.2017ZX10104001,2018ZX10711001,and 2018ZX10713002]
文摘Enteroviruses (EVs) are members of the genus Enteroviruswithin the orderPicornavirales, family Picornaviridae, and consist of 12 species, including EV-A, EV-B, EV-C, and EV-D, which are associated with human infections. Coxsackievirus B6 (CV-B6)belongs to the species EV-B, which currently consists of 63 serotypes, including echovirus (serotypes 1–7,9, 11–21, 24–27, 29–33), coxsackievirus group A (CVA9), coxsackievirus group B (CV-B, serotypes 1–6),the newly identified EVs (serotypes EV-B69, B73–75,B77–88, B93, B97–98, B100–101, B106–107, and B110–113), and the simian enterovirus SA5.
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).
基金the National Key Research and Development Program of China(2016YFD0500301)the National Major Project for Control and Prevention of Infectious Disease in China(2018ZX10101002 and 2017YFC1200503)+1 种基金the Bureau of Science and Information Technology of Guangzhou Municipality,China(201604020011,201704020219)National Key R&D Program of China(2018ZX10732401).
文摘Zika virus(ZIKV) causes rash, moderate fever, conjunctivitis, and arthralgia, and has serious connection with neurological complications;therefore, it is a major threat to public health. A rapid and supersensitive method for detecting anti-ZIKV antibodies in humans and animals is thus urgently required. Here, we report an NS1-based luciferase immunosorbent assay(LISA), developed to detect ZIKV-specific IgG. Fusion proteins including a reporter Nano-luciferase(NLuc) and various fragments of ZIKV NS1 protein were expressed in 293 T cells. LISA was performed using the above cell lysates containing the expressed fusion proteins. Sample panels of humans and animals infected with ZIKV were examined for sensitivity of LISA, relative to those of ZIKV RT-PCR, commercial NS1-based ELISA, and micro-neutralization(MN) assays.Specificity and potential cross-reactivity were also evaluated using various convalescent serum samples derived from patients infected with dengue virus(DENV), Japanese encephalitis virus(JEV), and hepatitis C virus(HCV). Results indicated the optimal antigenic domain for anti-ZIKV IgG detection was located within 172–352 amino acids(aa) of ZIKV NS1 protein. NS1-based LISA performs better than commercial ELISA in anti-ZIKV Ig G detection. LISA was shown to be at least fourfold more sensitive than commercial ELISA, and could detect anti-ZIKV Ig G in various animal hosts without the need of species-specific labeled antibody. This novel assay is potentially useful for the rapid and sensitive detection of anti-ZIKV IgG in human and animal samples.
基金supported by the National Science and Technology Major Project [2018ZX10713-002,2017ZX10104001,2018ZX10711001]National key research and development project [2017YFC1200505,2016YFC1200905]the Development Grant of State Key Laboratory of Infectious Disease Prevention and Control [2015SKLID505,2014SKLID103]
文摘Objective The current outbreak of Zika virus(ZIKV)poses a severe threat to human health.Two ZIKV strains were isolated from mosquitoes collected from the Dejiang prefecture in China in 2016,which was the first isolation of ZIKV in nature in China.Methods In this study,serum samples were collected from 366 healthy individuals and 104 animals from Dejiang prefecture in 2017,and the plaque reduction neutralization test(PRNT)was used to evaluate the seroprevalence of ZIKV.Results None of the 366 residents from whom the samples were collected were seropositive for ZIKV.None of the 11 pigs from whom the samples were collected were seropositive for ZIKV,while 1 of 63(1.59%)chickens and 2 of 30(6.67%)sheep were seropositive for ZIKV.Conclusions The extremely low seropositivity rate of ZIKV antibodies in animals in the Dejiang prefecture,Guizhou province in this study indicates that ZIKV can infect animals;however,there is a low risk of ZIKV circulating in the local population.
基金supported by the National Key Research and Development Project of China(2016YFD0500304)the National Science and Technology Major Project of China(2018ZX09711003)+3 种基金the National Natural Science Foundation of China(81621005 and 31770190)supported by the National Science Fund for Distinguished Young Scholars(81925025)the Innovative Research Group(81621005)from the NSFCthe Innovation Fund for Medical Sciences(2019-I2M-5–049)from the Chinese Academy of Medical Sciences
文摘West Nile virus(WNV)is an important neurotropic flavivirus that is widely distributed globally.WNV strain XJ11129 was first isolated in Xinjiang,China,and its genetic and biological characteristics remain largely unknown.In this study,phylogenetic and sequence analyses revealed that XJ11129 belongs to lineage 1 a and shares high genetic identity with the highly pathogenic strain NY99.Then,the full-length genomic c DNA of XJ11129 was amplified and assembled using a modified Gibson assembly(GA)method.The virus(named r XJ11129)was successfully rescued in days following this method.Compared with other wild-type WNV isolates,r XJ11129 exhibited virulence indistinguishable from that of the NY99 strain in vivo.In summary,the genomic and virulence phenotypes of r XJ11129 were characterized in vivo and in vitro,and these data will improve the understanding of the spread and pathogenesis of this reemerging virus.
基金supported by the National Key Program for Infectious Disease of China(2018ZX10101002)the National Key Research and Development Program of China(2016YFD0500301,2020YFC0840900)。
文摘Dear Editor,A novel coronavirus disease(COVID-19)caused by acute respiratory syndrome coronavirus 2(SARS-Co V-2)first broke out in Wuhan,Hubei Province,China,in 2019.More than 50 million cases of COVID-19 have been confirmed globally with 1,280,868 deaths reported as of November12,2020(https://covid19.who.int/).
基金the National Key Research and Development Program of China(2021YFC0863400,2022YFC2604105).
文摘Emerging and re-emerging viruses from wild animals have seriously threatened the health of humans and domesticated animals in recent years.Herein,we isolated a new mammalian orthoreovirus(MRV),Pika/MRV/GCCDC7/2019(PMRV-GCCDC7),in the Qinghai-Tibet Plateau wild pika(Ochotona curzoniae).Though the PMRVGCCDC7 shows features of a typical reovirus with ten gene segments arranged in 3:3:4 in length,the virus belongs to an independent evolutionary branch compared to other MRVs based on phylogenetic tree analysis.The results of cellular susceptibility,species tropism,and replication kinetics of PMRV-GCCDC7 indicated the virus could infect four human cell lines(A549,Huh7,HCT,and LoVo)and six non-human cell lines,including Vero-E6,LLCMK2,BHK-21,N2a,MDCK,and RfKT cell,derived from diverse mammals,i.e.monkey,mice,canine and bat,which revealed the potential of PMRV-GCCDC7 to infect a variety of hosts.Infection of BALB/c mice with PMRVGCCDC7 via intranasal inoculation led to relative weight loss,lung tissue damage and inflammation with the increase of virus titer,but no serious respiratory symptoms and death occurred.The characterization of the new reovirus from a plateau-based wild animal has expanded our knowledge of the host range of MRV and provided insight into its risk of trans-species transmission and zoonotic diseases.
基金National Key Research and Development Program of China,Grant/Award Number:2021YFC2301403 and 2022YFF0711000。
文摘Background:This study aimed to construct and characterize a humanized influenza mouse model expressing hST6GAL1.Methods:Humanized fragments,consisting of the endothelial cell-specific K18 promoter,human ST6GAL1-encoding gene,and luciferase gene,were microinjected into the fertilized eggs of mice.The manipulated embryos were transferred into the oviducts of pseudopregnant female mice.The offspring were identified using PCR.Mice exhibiting elevated expression of the hST6GAL1 gene were selectively bred for propagation,and in vivo analysis was performed for screening.Expression of the humanized gene was tested by performing immunohistochemical(IHC)analysis.Hematologic and biochemical analyses using the whole blood and serum of humanized hST6GAL1 mice were performed.Results:Successful integration of the human ST6GAL1 gene into the mouse genome led to the overexpression of human SiaT ST6GAL1.Seven mice were identified as carrying copies of the humanized gene,and the in vivo analysis indicated that hST6GAL1gene expression in positive mice mirrored influenza virus infection characteristics.The IHC results revealed that hST6GAL1 was expressed in the lungs of humanized mice.Moreover,the hematologic and biochemical parameters of the positive mice were within the normal range.Conclusion:A humanized influenza mouse model expressing the hST6GAL1 gene was successfully established and characterized.
文摘TheRhabdoviridae family comprises a diverse range of negative-sense single-stranded ribonucleic acid(RNA)viruses,including significant human and mammalian viruses transmitted by various arthropod species.Herein,usingAedes albopictus(Ae.albopictus)samples collected in two urban parks during 2023 and 2024,through metagenomics sequencing,16 sequences were identified as putative novel viruses,showing closest homology to insect-specific viruses,mycoviruses,or plant-associated viruses.Notably,two novel viruses,Aedes albopictus almendravirus GCCDC15(Aealb-AlmV GCCDC15)and Aedes albopictus almendravirus GCCDC16(Aealb-AlmV GCCDC16)were identified and successfully isolated.Both of these viruses belong to the genusAlmendravirus within theRhabdoviridae family.Phylogenetic analysis revealed that Aealb-AlmV GCCDC15 and GCCDC16 are distantly related to Coot Bay virus(the United States of America,2013)and Menghai rhabdovirus(Yunnan Province,China,2017).The genetic distances between these two viruses and their most similar viruses are marked by 59.85%and 87.20%of amino acid identity in the L protein,respectively,supporting their classification as two new species in theRhabdoviridae family.Cytopathic effects and rod-like virions were observed in mosquito cells(C6/36)after inoculating with supernatants from theAe.albopictus samples.To investigate the natural distribution and persistence of the novel almendraviruses,we conducted a specific reverse transcription-polymerase chain reaction(RT-PCR)screening ofAe.albopictus mosquitoes collected from two urban parks across different time points.The assays confirmed the presence of both Aealb-AlmV GCCDC15 and GCCDC16 in mosquito populations.Critically,these viruses were detected repeatedly over successive sampling periods and in mosquitoes from geographically distinct sites within the urban environment.In summary,our study delineates the virome characteristics ofAedes mosquitoes in the urban ecosystem and successfully isolated two novel rhabdoviruses.The recurrent detection provides clear evidence for the sustained circulation ofAe.albopictus-derived almendraviruses in urban parks,highlighting their ongoing transmission and establishment in these habitats.
