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Diagnostic assays developed for the control of foot-andmouth disease in India

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World Journal of Virology 2015年第3期4卷 295-302页

作者： Gaurav Kumar Sharma Sonalika Mahajan Rakesh Matura Saravanan Subramaniam Rajeev Ranjan Jitendra Biswal Manoranjan Rout Jajati Keshari Mohapatra Bana Bihari Dash Aniket Sanyal Bramhadev Pattnaik project directorate on foot and mouth disease Indian Council of Agricultural Research

foot-and-mouth disease(FMD) is a highly contagious and economically devastating disease of livestock, primarily affecting cattle, buffalo and pigs. FMD virus serotypes O, A and Asia1 are prevalent in India and systematic efforts are on to control and eventually eradicate the disease from the country. FMD epidemiology is complex due to factors like co-circulation, extinction, emergence and re-emergence of genotypes/lineages within the three serotypes, animal movement, diverse farm practices and large number of susceptible livestock in the country. Systematic vaccination, prompt diagnosis, strict biosecurity measures, and regular monitoring of vaccinal immunity and surveillance of virus circulation are indispensible features for the effective implementation of the control measures. Availability of suitable companion diagnostic tests is very important in this endeavour. In this review, the diagnostic assays developed and validated in India and their contribution in FMD control programme is presented.

关键词： foot-and-mouth disease Diagnosis Serosurveillance Sero-monitoring Multiplex polymerase chain reaction Real-time polymerase chain reaction Lineage differentiating polymerase chain reaction

African Swine Fever Virus MGF-110-9L-deficient Mutant Has Attenuated Virulence in Pigs

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Virologica Sinica 2021年第2期36卷 187-195页

作者： Dan Li Yinguang Liu Xiaolan Qi Yuan Wen Pan Li Zhao Ma Yongjie Liu Haixue Zheng Zhijie Liu State Key Laboratory of Veterinary Etiological Biology and OIE/National foot and mouth disease Reference Laboratory Lanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhou 730046China

African swine fever virus(ASFV)is the etiological agent of African swine fever(ASF),an often lethal disease in domestic and wild *** represents a major threat to the swine industry ***,no commercial vaccine is available because of the complexity of ASFV or biosecurity *** attenuated viruses that are naturally isolated or genetically manipulated have demonstrated reliable protection against homologous ASFV strain *** the present study,a mutant ASFV strain with the deletion of ASFV MGF-110-9L(ASFV-D9L)was generated from a highly virulent ASFV CN/GS/2018 parental strain,a genotypeⅡ*** to the parental ASFV isolate,deletion of the MGF-110-9L gene significantly decreased the ability of ASFV-D9L to replicate in vitro in primary swine macrophage cell *** majority of animals inoculated intramuscularly with a low dose of ASFV-D9L(10 HAD50)remained clinically normal during the 21-day observational *** of five ASFV-D9L-infected animals displayed low viremia titers and low virus shedding and developed a strong virus-specific antibody response,indicating partial attenuation of the ASFV-D9L strain in *** findings imply the potential usefulness of the ASFV-D9L strain for further development of ASF control measures.

关键词： African swine fever virus(ASFV) MGF-110-9L Mutant Attenuated virulence Pig

Preparation and immunoprotective effects of a virus-like particle candidate vaccine of the dominant epidemic D3 genotype coxsackievirus A6 in China

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Biosafety and Health 2024年第1期6卷 28-34页

作者： Xiaoliang Li Xizhu Xu Jichen Li Huanhuan Lu Congcong Wang Rui Wang Jinbo Xiao Ying Liu Yang Song Jingdong Song Qiang Sun Yong Zhang Department of Public Health and Health Administration Shandong First Medical UniversityJinan 250117China National Key Laboratory of Intelligent Tracking and Forecasting for Infectious diseases(NITFID) National Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China National Polio Laboratory WHO WPRO Regional Polio Reference LaboratoryNational Health Commission Key Laboratory for BiosafetyNational Health Commission Key Laboratory for Medical VirologyNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBejing 102206China Department of Medical Microbiology Weifang Medical UniversityWeifang 261053China

Coxsackievirus A6 of the D3a genotype(CVA6 D3a)is a primary pathogen causingmainland of China's hand,foot,and mouth disease(HFMD).Viral‐like particle(VLP)vaccines represent a potential candidate vaccine to prevent *** study collected Anti‐CVA6 D3a VLPs serum from BALB/c female mice immunized using CVA6 D3a *** neutralizing antibody levels were compared against the representative 14‐JX2018(D3a)and N4‐YN2015(D3b)strains between the antisera of different immune *** immunoprotective effect of anti‐CVA6 D3a VLPs against these strains was monitored using pathological sections and immuno-histochemical results of lung and skeletal muscle tissues in seven‐day‐old Institute of Cancer Research(ICR)*** protection against different branches of viruses was evaluated in 7‐day‐old(serum passive immune protection)and 14‐day‐old(VLPs active immune protection)neonatal ICR mice ***‐neutralizing antibody levels were positively correlated with the number of immunizations and higher against 14‐JX2018 than against N4‐***,these levels were significantly higher with abdominal injection than intramuscular *** immunized serum of 7‐day‐old ICR mice inoculated three times was 100%protected against CVA6 D3a 14‐JX2018(lethal titer:106.25 TCID 50)and CVA6 D3b N4‐YN2015(lethal titer:105.25TCID 50)fatal attacks,*** ICR mice that have completed two active immunizations for 14 days,both CVA6 D3a 14‐JX2015(challenge titer:108.25 TCID 50)and CVA6 D3b N4‐YN2015(challenge titer:107.25 TCID 50)were used for the challenge,and the mice were able to ***,the CVA6 D3a VLPs prepared in this study are a potential vaccine candidate for CVA6,as it has the optimal protective effect against both CVA6 D3a and D3b evolutionary branches viruses.

关键词： Coxsackievirus A6 D3 genotype Virus‐like particle Immunoprotective effect

Coxsackievirus B3 HFMD animal models in Syrian hamster and rhesus monkey

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Virologica Sinica 2024年第2期39卷 290-300页

作者： Suqin Duan Wei Zhang Yongjie Li Yanyan Li Yuan Zhao Weihua Jin Quan Liu Mingxue Li Wenting Sun Lixiong Chen Hongjie Xu Jie Tang Jinghan Hou Zijun Deng Fengmei Yang Shaohui Ma Zhanlong He Institute of Medical Biology Chinese Academy of Medical Sciences Medical Primate Research Center Peking Union Medical CollegeYunnan Key Laboratory of Vaccine Research Development on Severe Infectious DiseaseKunming650118China

Coxsackievirus B3(CVB3)is the pathogen causing hand,foot and mouth disease(HFMD),which manifests across a spectrum of clinical severity from mild to ***,CVB3-infected mouse models mainly demonstrate viral myocarditis and pancreatitis,failing to replicate human HFMD *** several enteroviruses have been evaluated in Syrian hamsters and rhesus monkeys,there is no comprehensive data on *** this study,we have first tested the susceptibility of Syrian hamsters to CVB3 infection via different *** results showed that Syrian hamsters were successfully infected with CVB3 by intraperitoneal injection or nasal drip,leading to nasopharyngeal colonization,acute severe pathological injury,and typical HFMD ***,the nasal drip group exhibited a longer viral excretion cycle and more severe pathological *** the subsequent study,rhesus monkeys infected with CVB3 through nasal drips also presented signs of HFMD symptoms,viral excretion,serum antibody conversion,viral nucleic acids and antigens,and the specific organ damages,particularly in the ***,there were no significant differences in myocardial enzyme levels,and the clinical symptoms resembled those often associated with common,mild *** summary,the study successfully developed severe Syrian hamsters and mild rhesus monkey models for CVB3-induced *** models could serve as a basis for understanding the disease pathogenesis,conducting pre-trial prevention and evaluation,and implementing post-exposure intervention.

关键词： Coxsackievirus B3(CVB3) Hand foot and mouth disease(HFMD) Animal models Syrian hamster Rhesus monkey

Expression of Major B-cell Epitopes within the 2C Non-structural Protein of FMDV and Their Immunoreactivity

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畜牧兽医学报 2009年第S1期40卷 39-42页

作者： FU Yuan-fang LU Zeng-jun TIAN Mei-na ZHANG Xiao-li LIU Zai-xin CAI Xue-peng State Key Laboratory of Veterinary Etiological Biology National Foot-and-Mouth Disease Reference LaboratoryLanzhou Veterinary Research InstituteChinese Academy of Agricultural Sciences

A pair of specific primers was designed from the 2C gene sequence of foot-and-mouth disease virus(FMDV)for amplification of a fragment including 174bp of the 5'-end and 279bp of the 3'-end of the2C gene,which encoded an abundance of known B-cell epitopes of the *** amplified fragment was inserted into pET-30a plasmid(Novagen)via two unique endonuclease restriction sites,Nco I and Sal *** confirmed that the open reading frame of interest was correctly inserted into the positive recombinant *** positive plasmid was transformed into the host bacteria BL21(DE3)pLys for protein *** induction by IPTG at 37℃for 5 hours,the expressed product was analyzed by SDSPAGE and Western blotting,confirming successful *** product is a 23kDa fusion protein and was shown to react with sera derived from FMDV-infected *** approach provides an useful antigen for establishing an enzyme-linked immunoelectro-transfer blot assay(EITB)diagnostic method,useful for differentiating FMDV-infected animals from those that been vaccinated.

关键词： FMDV nonstructural protein 2C gene EITB

The Distribution of Different Clades of Seneca Valley Viruses in Guangdong Province,China

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Virologica Sinica 2018年第5期33卷 394-401页

作者： Pan Chen Fan Yang Weijun Cao Huanan Liu Keshan Zhang Xiangtao Liu Zhiwen Xu Zixiang Zhu Haixue Zheng State Key Laboratory of Veterinary Etiological Biology National Foot and Mouth Diseases Reference LaboratoryKey Laboratory of Animal Virology of Ministry of Agriculture Lanzhou Veterinary Research Institute Chinese Academy of Agricultural Sciences Animal Biotechnology Center Key Laboratory of Animal Disease and Human Health of Sichuan Province Sichuan Agricultural University

Seneca Valley virus (SVV), a newly determined etiological agent of vesicular disease in swine, causes porcine idiopathic disease and occasional acute death in piglets. Recently, an increased number of SVV infection cases have been reported in the United States (US) and China, resulting in significant economic losses to the swine industry. The first identification of SVV in China was reported in Guangdong Province, a major swine producing province. The cases of SVV were continuously reported in Guangdong in 2015 and 2016. However, the spread of SVV in Guangdong in 2017 remains *** this study, we determined two new SVV strains, CH-GD-2017-1 and CH-GD-2017-2, from Guangdong. The genetic analysis suggested that the two Guangdong strains showed different characteristics to previous Guangdong strains. They showed lower nucleotide similarity with strains isolated in 2015 and 2016, and were more similar to the US *** analyses indicated that the new strains were clustered in a different clade with previous Guangdong *** found 28 mutated amino acids in the new strains, compared with the first Guangdong strain, SVV CH-01-2015. In the geographic analysis, we found that the US and China reported more SVV cases than other countries, and most of the SVV cases were reported in east and central China—of which, Guangdong Province is one of the major epidemic regions. In conclusion, our findings indicate that SVV continued to spread in Guangdong Province in 2017, and two different clades of SVVs have emerged in this region.

关键词： Seneca Valley virus (SVV) Genomic sequence Phylogenetic tree Epidemiological analysis

foot-and-mouth disease Virus Inhibits RIP2 Protein Expression to Promote Viral Replication

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Virologica Sinica 2021年第4期36卷 608-622页

作者： Huisheng Liu Qiao Xue Zixiang Zhu Fan Yang Weijun Cao Xiangtao Liu Haixue Zheng State Key Laboratory of Veterinary Etiological Biology National Foot and Mouth Diseases Reference LaboratoryKey Laboratory of Animal Virology of Ministry of AgricultureLanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhou 730046China

Receptors interaction protein 2(RIP2)is a specific adaptor molecule in the downstream of *** role of RIP2 during foot-and-mouth disease virus(FMDV)infection remains ***,our results showed that RIP2 inhibited FMDV replication and played an important role in the activation of IFN-βand NF-κB signal pathways during FMDV *** infection triggered RIP2 transcription,while it reduced the expression of RIP2 *** analysis showed that FMDV 2B,2C,3C^(pro),and L^(pro) proteins were responsible for inducing the reduction of RIP2 protein.3C^(pro) and L^(pro) are viral proteinases that can induce the cleavage or reduction of many host proteins and block host protein *** carboxyl terminal 105-C114 and 135-C144 regions of 2B were essential for reduction of *** results also showed that the N terminal 1-61 region of 2C were essential for the reduction of *** 2C-induced reduction of RIP2 was dependent on inducing the reduction of poly(A)-binding protein 1(PABPC1).The interaction between RIP2 and 2C was observed in the context of viral infection,and the residues 1-61 were required for the *** data clarify novel mechanisms of reduction of RIP2 mediated by FMDV.

关键词： foot-and-mouth disease virus(FMDV) Receptors interaction protein 2(RIP2) PABPC1 2C Immune evasion

Establishment of an indirect immunofluorescence assay for the detection of African swine fever virus antibodies

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Journal of Integrative Agriculture 2024年第1期23卷 228-238页

作者： Wan Wang Zhenjiang Zhang Weldu Tesfagaber Jiwen Zhang Fang Li Encheng Sun Lijie Tang Zhigao Bu Yuanmao Zhu Dongming Zhao State Key Laboratory for Animal disease Control and Prevention National African Swine Fever Para-reference LaboratoryNational High Containment Facilities for Animal Diseases Control and PreventionHarbin Veterinary Research InstituteChinese Academy of Agricultural SciencesHarbin 150069China College of Veterinary Medicine Northeast Agricultural UniversityHarbin 150069China

African swine fever(ASF)continues to cause enormous economic loss to the global pig *** there is no safe and effective vaccine,accurate and timely diagnosis of ASF is essential to implement control *** immunofluorescence assay(IFA)is a gold standard serological method recommended by the World Organization for Animal Health(WOAH).In this study,we used primary fetal kidney cells to establish a wild boar cell line(BK2258)that supported the efficient replication of ASF virus(ASFV)SD/DY-I/21 and showed visible cytopathic effect(CPE).Moreover,using BK2258,we established a sensitive and specific IFA for ASFV antibody *** standardize and evaluate the performance of this assay,we used serum samples from pigs infected with the low virulent genotype I SD/DY-I/21 and genotype II HLJ/HRB1/20,and immunized with the vaccine candidate HLJ/18-7GD,field samples,and negative serum *** IFA reacted with the ASFV-positive sera and displayed bright fluorescence *** was no non-specific green fluorescence due to cellular senescence or other cell damage-causing *** to a commercial indirect enzyme-linked immunosorbent assay(iELISA),ASFV antibodies were detected 1–4 days earlier using our *** detection limits of the IFA and iELISA for the same ASFV-antibody positive serum samples were 1:25,600 and 1:6,400,respectively,indicating that the IFA is more sensitive than *** newly established IFA was highly specific and did not cross-react with sera positive for six other important porcine pathogens(i.e.,Classical swine fever virus(CSFV),Porcine reproductive and respiratory syndrome virus(PRRSV),Porcme circovirus type 2(PCV2),Pseudorabies virus(PRV),foot-and-mouth disease virus type O(FMDV/O),and Porcine epidemic diarrhea virus(PEDV)).This study thus provides a sensitive,specific,and reliable detection method that is suitable for the serological diagnosis of ASF.

关键词： African swine fever antibody IFA serological method

Porcine Picornavirus 3C Protease Degrades PRDX6 to Impair PRDX6-mediated Antiviral Function

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Virologica Sinica 2021年第6期36卷 948-957页

作者： Congcong Wang Huanhuan Feng Xiangle Zhang Kangli Li Fan Yang Weijun Cao Huisheng Liu Lili Gao Zhaoning Xue Xiangtao Liu Zixiang Zhu Haixue Zheng State Key Laboratory of Veterinary Etiological Biology OIE/National Foot and Mouth Diseases Reference LaboratoryKey Laboratory of Animal Virology of Ministry of AgricultureLanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhou 730046China

Peroxiredoxin-6(PRDX6)is an antioxidant enzyme with both the activities of peroxidase and phospholipase A2(PLA2),which is involved in regulation of many cellular ***,the function of PRDX6 during virus infection remains *** this study,we found that the abundance of PRDX6 protein was dramatically decreased in foot-and-mouth disease virus(FMDV)infected *** of PRDX6 inhibited FMDV *** contrast,knockdown of PRDX6 expression promoted FMDV replication,suggesting an antiviral role of *** explore whether the activity of peroxidase and PLA2 was associated with PRDX6-mediated antiviral function,a specific inhibitor of PLA2(MJ33)and a specific inhibitor of peroxidase activity(mercaptosuccinate)were used to treat the cells before FMDV *** results showed that incubation of MJ33 but not mercaptosuccinate promoted FMDV ***,overexpression of PRDX6 slightly enhanced type I interferon *** further determined that the viral 3Cprowas responsible for degradation of PRDX6,and 3Cpro-induced reduction of PRDX6 was independent of the proteasome,lysosome,and caspase *** protease activity of 3Cprowas required for induction of PRDX6 ***,PRDX6 suppressed the replication of another porcine picornavirus Senecavirus A(SVA),and the 3Cproof SVA induced the reduction of PRDX6 through its proteolytic activity as ***,our results suggested that PRDX6 plays an important antiviral role during porcine picornavirus infection,and the viral 3Cproinduces the degradation of PRDX6 to overcome PRDX6-mediated antiviral function.

关键词： Porcine picornavirus Peroxiredoxin-6(PRDX6) 3Cpro Phospholipase A2(PLA2) Antagonism