Percepción del riesgo biológico en el proceso productivo de la vacuna contra la peste porcina clásica / Biological risk perception in the production process of classical swine fever vaccine

La existencia de agentes biológicos como el virus de la peste porcina clásica para la producción de vacunas veterinarias, entre otros de igual importancia para nuestro país y la región en general, justifica una buena gestión de la seguridad biológica, ya que el desconocimiento del riesgo por parte del personal que labora en estas vacunas puede provocar contaminaciones de graves consecuencias medio ambientales, en el proceso de producción y a nivel personal si son causantes de accidentes fatales. El objetivo de la investigación fue realizar un análisis de la percepción de riesgo existente en el personal responsable del proceso de producción de la vacuna contra la peste porcina clásica. La aplicación del RISKPERCEP en el personal de la instalación de producción de la vacuna de la peste porcina clásica mostró como resultados el comportamiento de diferentes variables que hacen evidente la alta subestimación del riesgo existente en el personal evaluado y que existe la necesidad de profundizar en la formación en bioseguridad para todo el personal que labora en el proceso. Finalmente, se relacionan estos temas y su importancia para mejorar la calidad de la producción en estos procesos, así como incrementar el conocimiento acerca del riesgo biológico a todos los niveles(AU)

The existence of high-risk biological agents such as the classical swine fever virus for the production of veterinary vaccines, among others of equal importance for our country and the region in general, justifies good management of biological safety, since ignorance of the risk on part of the personnel who work in them, can cause contamination with serious consequences both at personal and environmental level, causing fatal accidents. The objective of the research was to carry out an analysis of the perception of existing risk in the personnel responsible for the production process of the vaccine against classical swine fever. The application of RISKPERCEP in the classical swine fever vaccine production facility showed as results the behavior of different variables that make evident the high underestimation of the existing risk in the evaluated personnel and that there is a need to deepen the training in biosafety to all staff working in the process. Finally, these issues and their importance to improve the quality of production in these processes are related, as well as to increase knowledge about biological risk at all levels(AU)

The potential efficacy of the E2-subunit vaccine to protect pigs against different genotypes of classical swine fever virus circulating in Vietnam

Histopathological Evaluation of the Efficacy for Plant-produced E2 Protein Vaccine against Classical Swine Fever Virus (CSFV) in Piglets

Classical swine fever (CSF), previously known as hog cholera, remains one of the most important swine-related contagious diseases worldwide. In order to eradicate classical swine fever virus (CSFV), it is commonly used in LOM-850 strain as a live attenuated CSF vaccine. However, there are symptoms of vaccination, such as the depression of feed intake, and difficulty of differentiation between infected and vaccinated hosts is impossible based on the antibodies induced. Nicotiana benthamiana were considered as an alternative to the production of recombinant vaccines on account of higher yields and levels of soluble protein than other models and crops in protein recombinant products. This study was conducted to evaluate histopathological validation of the plant-produced E2 fusion protein (ppE2) in piglets. The piglets were challenged by an injection of YC11WB strain in 7 days, 11 days and 14 days after one shot of the vaccination. The histopathological examination indicated that ppE2 can protect against lethal CSFV challenge at least 11 days of vaccination in piglets. These data suggest that the ppE2 can be an effective vaccine against CSFV in piglets.

Generation and characterization of recombinant classical swine fever virus C-strain expressing the cap protein of porcine circovirus type 2 / 生物工程学报

Classical swine fever (CSF), caused by classical swine fever virus (CSFV), is a devastating viral disease in swine, leading to significant economic losses to the pig husbandry. C-strain is one of the best modified live vaccines against CSF. The vaccine is highly safe and efficacious and can provide rapid and complete protection against essentially all genotypes of CSFV. Co-infections of pigs with CSFV and porcine circovirus type 2 (PCV2) occur frequently in the field, making it difficult to control the associated diseases. Here, a recombinant C-strain rHCLV-Cap expressing the Cap protein of PCV2 was constructed and evaluated in vitro and in vivo. The recombinant had comparable phenotypes to C-strain in cell cultures and rabbits. At ten days post-immunization, anti-E2, but not anti-Cap, antibodies were detected in the rabbits inoculated with the recombinant virus. Our study warrants further work to construct C-strain-based bivalent vaccines.

Bovine viral diarrhea virus infections in pigs: why is this situation important for Brazilian herds? / A infecção pelo vírus da diarreia viral bovina em suínos: por que essa situação é importante para os rebanhos brasileiros?

Swine can be infected by the bovine viral diarrhea virus (BVDV) under natural conditions. For this reason, further information and divulgation are needed regarding the pathogenicity of this virus in swine. This infection is practically unknown in the realm of pig farming, and, as it shares some similarities with the classical swine fever virus (CSFV), its diagnosis becomes a challenge for official sanitary programs. Studies have shown the absence of clinical signs in piglets and reproductive problems in sows due to BVDV infections. There is little research on the prevalence, risk factors, preventive measures and control of BVDV in pigs around the world. And in Brazil, the data is practically non-existent. At the time of diagnosis, comparing the most efficient laboratory tests such as virus neutralization, ELISA, RT-PCR, and immunofluorescence so as to minimize the risk of cross serological reactions when dealing with a persistent or transient infection, can be an important tool. Moreover, the practical implications for CSFV eradication programs are a main reason for the development of further research against this infection. Therefore, this paper aims to review various aspects of BVDV infection in pigs, and how this information can be important for Brazilian herds.(AU)

O suíno pode ser infectado pelo vírus da diarreia viral bovina (BVDV) em condições naturais, por isso são necessárias maiores informações e mais divulgação sobre a ação deste vírus nos suínos. Esta infecção é praticamente desconhecida na suinocultura e, devido a algumas semelhanças com vírus da peste suína clássica (VPSC), torna-se um desafio para os programas sanitários oficiais. Estudos revelam a ausência de sinais clínicos em leitões ao mesmo tempo em que evidenciam problemas reprodutivos em porcas devido à infecção do BVDV. Poucas são as pesquisas sobre a prevalência, fatores de riscos, medidas de prevenção e controle do BVDV em suínos no mundo e, no Brasil, os dados são praticamente inexistentes. No diagnóstico, comparar os exames laboratoriais mais eficientes como a virusneutralização, ELISA, RT-PCR e imunofluorescência, diante de uma infecção persistente ou transitória, e assim minimizar o risco de reações sorológicas cruzadas pode ser uma ferramenta fundamental. Ademais, as implicações práticas em programas de erradicação da PSC são um grande motivo para o desenvolvimento de mais pesquisas frente a esta infecção. Portanto, este trabalho pretende revisar diversos aspectos da infecção do BVDV em suínos evidenciando o quanto essa situação pode ser importante para os rebanhos brasileiros.(AU)

Antigenic characterization of classical swine fever virus YC11WB isolates from wild boar

Classical swine fever (CSF), a highly contagious disease that affects domestic pigs and wild boar, has serious economic implications. The present study examined the virulence and transmission of CSF virus strain YC11WB (isolated from a wild boar in 2011) in breeding wild boar. Virulence of strain YC11WB in domestic pigs was also examined. Based on the severe clinical signs and high mortality observed among breeding wild boar, the pathogenicity of strain YC11WB resembled that of typical acute CSF. Surprisingly, in contrast to strain SW03 (isolated from breeding pigs in 2003), strain YC11WB showed both acute and strong virulence in breeding pigs. None of three specific monoclonal antibodies (7F2, 7F83, and 6F65) raised against the B/C domain of the SW03 E2 protein bound to the B/C domain of strain YC11WB due to amino acid mutations (⁷²⁰K→R and ⁷²³N→S) in the YC11WB E2 protein. Although strains YC11WB and SW03 belong to subgroup 2.1b, they had different mortality rates in breeding pigs. Thus, if breeding pigs have not developed protective immunity against CSF virus, they may be susceptible to strain YC11WB transmitted by wild boar, resulting in severe economic losses for the pig industry.

Expression and antiviral activity of a chimeric porcinized monoclonal antibody (cHQ06) against E2 protein of classical swine fever virus / 生物工程学报

Classical swine fever (CSF), one of OIE-listed diseases, is a highly contagious and economically important disease of pigs. Classical swine fever virus (CSFV) is the causative agent of CSF. The capsid (C) protein and the glycoproteins Erns, E1 and E2, are structural components of the virus. E2 is the most immunogenic protein of the CSFV glycoproteins, inducing neutralizing antibodies that provide protection against lethal CSFV challenge. In a previous study, we developed a murine MAb HQ06 against the E2 protein of CSFV. In this study, the variable region genes from HQ06 and constant regions gene of swine antibody are fused and cloned into the eukaryotic expression vectors to establish a cell line which can stably express a chimeric porcinized MAb (cHQ06) against E2 in CHO cell. The purified cHQ06 antibody protein was determined to be successfully generated, which exhibited high reactivity between cHQ06 and the E2 protein of CSFV by enzyme-linked immunosorbent assay (ELISA) and Western blotting. More importantly, we investigated the neutralizing activity of cHQ06 against CSFV. In conclusion, this study generated cHQ06 for efficient and stable production which can be used against to develop novel diagnostic assays, investigate the structure and function of the E2 protein and generate novel preparations of diagnosis and treatment.

Co-expression of the C-terminal domain of Yersinia enterocolitica invasin enhances the efficacy of classical swine-fever-vectored vaccine based on human adenovirus.

The use of adenovirus vector-based vaccines is a promising approach for generating antigen-specific immune responses. Improving vaccine potency is necessary in other approaches to address their inadequate protection for the majority of infectious diseases. This study is the first to reconstruct a recombinant replication-defective human adenovirus co-expressing E2 and invasin C-terminal (InvC) glycoproteins (rAd-E2-InvC). rAd-E2-InvC with 2×106 TCID50 was intramuscularly administered two times to CSFV-free pigs at 14 day intervals. No adverse clinical reactions were observed in any of the pigs after the vaccination. The CSFV E2-specific antibody titer was significantly higher in the rAd-E2-InvC group than that in the rAdV-E2 group as measured by NPLA and blocking ELISA. Pigs immunized with rAd-E2-InvC were completely protected against lethal challenge. Neither CSFV RNA nor pathological changes were detected in the tissues after CSFV challenge. These results demonstrate that rAd-E2-InvC could be an alternative to the existing CSF vaccine. Moreover, InvC that acts as an adjuvant could enhance the immunogenicity of rAdV-E2 and induce high CSFV E2-specific antibody titer and protection level.

Screening of cellular proteins that interact with the classical swine fever virus non-structural protein 5A by yeast two-hybrid analysis.

Classical swine fever virus (CSFV), the pathogen of classical swine fever (CSF), causes severe hemorrhagic fever and vascular necrosis in domestic pigs and wild boar. A large number of evidence has proven that non-structural 5A (NS5A) is not only a very important part of viral replication complex, but also can regulate host cell’s function; however, the underlying mechanisms remain poorly understood. In the current study, aiming to find more clues in understanding the molecular mechanisms of CSFV NS5A’s function, the yeast two-hybrid (Y2H) system was adopted to screen for CSFV NS5A interactive proteins in the cDNA library of the swine umbilical vein endothelial cell (SUVEC). Alignment with the NCBI database revealed 16 interactive proteins: DDX5, PSMC3, NAV1, PHF5A, GNB2L1, CSDE1, HSPA8, BRMS1, PPP2R3C, AIP, TMED10, POLR1C, TMEM70, METAP2, CHORDC1 and COPS6. These proteins are mostly related to gene transcription, protein folding, protein degradation and metabolism. The interactions detected by the Y2H system should be considered as preliminary results. Since identifying novel pathways and host targets, which play essential roles during infection, may provide potential targets for therapeutic development. The finding of proteins obtained from the SUVEC cDNA library that interact with the CSFV NS5A protein provide valuable information for better understanding the interactions between this viral protein and the host target proteins.

Production and characterization of monoclonal antibodies against a local isolate of classical swine fever virus.

Monoclonal antibodies (mAbs) against a classical swine fever virus (CSFV; subgenogroup 1:1) isolate from Assam, India were produced and characterized. Four fusions of myeloma cells (SP2/0Ag) were made with spleenocytes of 8-10 weeks old BALB/C mice immunized with the viral antigen. Several hybridoma clones secreting antibodies to the virus were obtained after four fusions, but five hybridoma clones secreting antibody specific to the virus could be stabilized. All the mAbs belong to the IgG2a isotype. Except one, none of the four mAbs showed cross reaction with bovine viral diarrhoea virus and border disease virus (BDV). One mAb showed cross reaction with BDV. All the four mAbs specific to CSFV showed reactivity with the parental virus in immunoperoxidase test (IPT) and with a single protein band (molecular weight 55 kD approximately) of the virus in western blotting. In neutralization peroxidase linked assay (NPLA) all the mAbs reacted with 13 CSFV local isolates as well as with the cell culture adapted lapinized vaccine virus strain belonging to the subgenogroup 1:1. This is the first report on production and characterization of mAbs against CSFV in India.

Development of Multiple ELISAs for the Detection of Antibodies against Classical Swine Fever Virus in Pig Sera / 中国病毒学·英文版

The major immunogenic proteins (Ems,E2 and NS3) of classical swine fever virus (CSFV) (Shimen strain) were expressed in E.coli and purified by affinity chromatography.The recombinant antigens were applied to develop multiple enzyme-linked immunosorbent assays (ELISAs) for the detection of specific antibodies in pig sera.Optimum cut-off values were determined by receiver operating characteristic (ROC) analysis after testing 201 sera of vaccinated pigs and 64 negative sera of unvaccinated piglets.The multiple ELISAs were validated with 265 pig sera yielding high sensitivity and specificity in comparison with the virus neutralization results.The results demonstrated that multiple ELISAs can be a valuable tool for the detection of CSFV infection and serological surveys in CSFV-free countries or for the evaluation of the antibody responses in pigs induced by a live attenuated C-strain vaccination.

Analysis of in vitro apoptosis induced by virulent Korean isolate of classical swine fever virus in peripheral blood B cell line / 대한수의학회지

Classical swine fever (CSF) is a highly contagious disease among swine that has an important economic impact on worldwide. One clinical symptom of CSF is leukopenia, in particular lymphopenia, which is a characteristic event that occurs early in the course of CSF. Though lymphopenia associated with apoptosis, the pathogenic mechanism underlying the lymphopenia has not been well studied. To understand these mechanisms, we investigated the response of porcine B cell lines to infection with SW03, virulent strain isolated from swine tissue in Korea. This study demonstrated that SW03-infected L35 cell were induced apoptosis through the detection of activated caspase-3. In addition, SW03 infection leaded to alterations in pro-apoptotic, Bax, and anti-apoptotic, Bcl-xL proteins of Bcl-2 family. Our results would suggest that SW03-infected L35 cells induced apoptosis via intrinsic mitochondrial pathway.

Establishment and characterization of an infectious cDNA clone of a classical swine fever virus LOM strain

Classical swine fever virus (CSFV) causes a highly contagious disease among swine that has an important economic impact worldwide. CSFV strain LOM is an attenuated virus of low virulent strain of Miyagi isolated from Japan in 1956. Eight DNA fragments representing the genome of the CSFV strain LOM were obtained by RT-PCR. These were used to determine the complete nucleotide sequence and construct a full-length cDNA clone which was called Flc-LOM. Sequence analysis of the recombinant clone (Flc-LOM) revealed the presence of eight mutations, resulting in two amino acid substitutions, when compared to the parental sequence. RNA transcripts of both LOM and Flc-LOM were directly infectious in PK-15 cells. The rescued Flc-LOM virus grew more slowly than the parental virus, LOM, in the cells. Intramuscular immunization with Flc-LOM was safe and highly immunogenic in pigs; no clinical signs or virus transmission to sentinel animals were observed after 35 days. CSFV-specific neutralizing antibodies were detected 14 days post-infection. After challenge with the virulent CSFV strain SW03, pigs immunized with Flc-LOM were shown to be fully protected. Thus, our newly established infectious clone of CSFV, Flc-LOM, could serve as a vaccine candidate.

A Novel RT-LAMP Assay for Rapid and Simple Detection of Classical Swine Fever Virus / 中国病毒学·英文版

A simple and rapid assay for the detection of Classical swine fever virus(CSFV)was established using reverse transcription loop-mediated isothermal amplification(RT-LAMP).This study describes the amplification of the genomic RNA of CSFV under isothermal conditions(63℃)within one hour,using a set of six primers(two outer primers,two inner primers and two loop primers).This RT-LAMP assay showed 100-fold higher sensitivity than the standard RT-PCR method and identified eighteen additional positive cases that were negative when tested by RT-PCR.This RT-LAMP was able to detect all the 13 strains of CSFV but not the BVDV.PRRSV.SIV.PRV-PCV,thus showed a good specificity.Products amplified by RT-LAMP can be visualized by agarose gel electrophoresis and in addition,either as a white precipitate at the bottom of the tube after a pulse spin or as a color change when dyed with SYBR Green I which are visible to the naked eye.Because RT-LAMP is low-cost and produces rapid results,it has the potential to be an excellent tool for CSFV surveillance in the field,especially in developing countries.

An Indirect ELISA of Classical Swine Fever Virus Based on Quadruple Antigenic Epitope Peptide Expressed in E.coli / 中国病毒学·英文版

In this study,a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV)E2 glycoprotein was expressed in E.coli to a obtain target protein.This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs.The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve(ROC)analysis based on 30 negative sera and 80 positive samples.The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination(IHA)test.The inter-assay and intra-assay coefficients of variation (CVs)for 16 sera were both ≤6.8%.No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus(BVDV)antibodies was observed.

Selection of a Less Pathogenic BVDV Strain for the Construction of Avirulent Chimeric Pestivirus

To select a less pathogenic bovine viral diarrhea virus (BVDV) strain for the construction of chimeric pestivirus harboring classical swine fever virus (CSFV) E2 gene, five Korean BVDV isolates (four type 1 isolates and a type 2 isolate) were evaluated for their pathological and biological properties with respect to porcine infection. Each of five groups of 100-day-old pigs was inoculated intranasally with one of the five BVDV isolates. No clinical sign or leukopenia was observed in any pig throughout the duration of the experiment, but viruses were detected in blood, nasal discharges and postmortem samples using RT-PCR. These results indicated that although the five BVD viruses could infect pigs, they did not cause clinically apparent symptoms. Because of its proper infection dynamics shown in this preliminary animal study and its fast growth rate and quick CPE in cell culture, one isolate (KD26-1) was chosen among the five isolates to test its virulence and immunogenic properties in 40-day-old piglets. Neither clinical sign nor pathological lesion was observed in 40-day-old piglets during the course of infection of isolate KD26-1. The first neutralizing antibodies were detectable 14 days post-inoculation (PI) and increased to 1:128~1:256 28 days PI. A BVDV specific gene was detectable by RT-PCR in tonsil, spleen, inguinal lymph node and brain until 14 days PI. According to this study, it can be concluded that isolate KD26-1 has little pathological effect in pigs and is a candidate for construction of chimeric pestivirus harboring CSFV E2 gene.

Selection of a Less Pathogenic BVDV Strain for the Construction of Avirulent Chimeric Pestivirus

To select a less pathogenic bovine viral diarrhea virus (BVDV) strain for the construction of chimeric pestivirus harboring classical swine fever virus (CSFV) E2 gene, five Korean BVDV isolates (four type 1 isolates and a type 2 isolate) were evaluated for their pathological and biological properties with respect to porcine infection. Each of five groups of 100-day-old pigs was inoculated intranasally with one of the five BVDV isolates. No clinical sign or leukopenia was observed in any pig throughout the duration of the experiment, but viruses were detected in blood, nasal discharges and postmortem samples using RT-PCR. These results indicated that although the five BVD viruses could infect pigs, they did not cause clinically apparent symptoms. Because of its proper infection dynamics shown in this preliminary animal study and its fast growth rate and quick CPE in cell culture, one isolate (KD26-1) was chosen among the five isolates to test its virulence and immunogenic properties in 40-day-old piglets. Neither clinical sign nor pathological lesion was observed in 40-day-old piglets during the course of infection of isolate KD26-1. The first neutralizing antibodies were detectable 14 days post-inoculation (PI) and increased to 1:128~1:256 28 days PI. A BVDV specific gene was detectable by RT-PCR in tonsil, spleen, inguinal lymph node and brain until 14 days PI. According to this study, it can be concluded that isolate KD26-1 has little pathological effect in pigs and is a candidate for construction of chimeric pestivirus harboring CSFV E2 gene.

Expression of Major Antigen Domains of Gene of E2 CSFV and Analysis of its Immunological Activity / 中国病毒学·英文版

E2 is an envelope glycoprotein of Classical swine fever virus (CSFV) and contains sequential neutralizing epitopes to induce virus-neutralizing antibodies and mount protective immunity in the natural host. In this study, four antigen domains (ABCD) of the E2 gene was cloned from CSFV Shimen strain into the retroviral vector pBABE puro and expressed in eukaryotic cell (PK15) by an retroviral gene expression system, and the activity of recombinant E2 protein to induce immune responses was evaluated in rabbits. The results indicated that recombinant E2 protein can be recognized by fluorescence antibodies of CSFV and CSFV positive serum (Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China) using Western blot, indirect immunofluorescence antibody test (IFAT) and ELISA, Furthermore, anti-CSFV specific antibodies and lymphocyte proliferation were elicited and increased by recombinant protein after vaccination. In the challenge test, all of rabbits vaccinated with recombinant protein and Chinese vaccine strain (C-strain) were fully protected from a rabbit spleen virus challenge. These results indicated that a retroviral-based epitope-vaccine carrying the major antigen domains of E2 is able to induce high level of epitope-specific antibodies and exhibits similar protective capability with that induced by the C-strain, and encourages further work towards the development of a vaccine against CSFV infection.

Classical Swine Fever in Brazil: study for the survey of classical swine fever outbreaks in Brazil from 1978 to 2004 / Peste Suína Clássica no Brasil: estudo para a avaliação dos surtos depeste suína clássica no Brasil de 1978 a 2004

The programs developed in Brazil with the aim to control and eradicate swine fever provided an opportunityfor the survey of Classical Swine Fever (CSF) outbreaks. Were concerned CSF official programs, strategies and results, during 26 years. Based in epizootic official data we showed that the number of CSF outbreaks from 1978 to 2004 drastically decreased in all country, although different eradicating strategies were applied in those official programs, especially in fourteen States of "CSF Free Zone". Were evaluated both CSF official programs: Swine Pests Combat Program (SPCP) from 1984 to 1991 and CSF Eradication and Control Program (CSFECP) from 1992 to 2004 by the decreasing of CSF outbreaks number. Considering the technical evolution in swine production systems, statistical analysis to compare the ranking of CSFoutbreaks in each program was performed by Mann-Whitney test, that showed at 95% confidence level(Table T) a significant difference (p< 0.05) between programs, as suggested in CSF outbreaks profileplotted diagram. The number of CSF outbreaks occurred from 2000­2004 in "CSF-infected" and "CSF-free" zones, was analyzed. Also, we regarded with most important recent CSF outbreak in Brazil occurred in 1997, during CSFECP, that was figured out by stamping out measures without appealing to preventive vaccination regimen. Those results suggest that the efficacy of implemented CSF eradication programs depends on the continuity of defined strategies as rigorous vigilance, notification, virus diagnostic screening and sanitary police measures in order to enable quick and adequate action upon CSFV detection

Os programas oficiais para o controle e erradicação de pestes suínas forneceram uma oportunidade de levantar o perfil de ocorrência da Peste Suína Clássica (PSC). Independente das estratégias aplicadasdurante 26 anos foi demonstrado que o número de surtos de PSC de 1978 até 2004 caiu drasticamente emtodo país, especialmente nos quatorze Estados inclusos na "Zona Livre de PSC". O estudo comparou o número de surtos de PSC durante a vigência do Programa de Combate às Pestes Suínas (PCPS) de 1984a 1991 e o Programa de Controle e Erradicação da PSC (PCEPSC) de 1992 a 2004. Considerando aevolução tecnológica nos sistemas de produção de suínos, a diferença nos resultados obtidos após aimplementação de cada programa foi avaliada pelo teste estatístico Mann Whitney por meio da ordenaçãodo número de surtos ocorridos. Essa análise demonstrou uma diferença significativa (p< 0,05) entre osprogramas no nível de confiança de 95% (Tabela T) com havia sido sugerido pelo diagrama do perfil deocorrência da PSC. A eficácia do PCEPSC para debelar o mais importante surto de PSC ocorridorecentemente no Brasil, em 1997, também foi considerada. Paralelamente, o número de surtos ocorridosde 2000 a 2004 nas áreas infectadas com a PSC e na zona livre de PSC foi avaliado. Os resultados sugeremque a eficácia dos programas de erradicação depende da continuidade das estratégias definidas como avigilância rigorosa, notificação, rastreamento do vírus e medidas sanitárias que agilizem a ação nomomento de detecção de vírus da PSC

Generation of a Recombinant Baculovirus Expressing The E2 Protein of Classical Swine Fever Virus and Its Immunogenicity in a Mouse Model / 生物化学与生物物理进展

Baculovirus-mediated gene transfer into mammalian cells has been used to develop non-replicative vector vaccines against a number of diseases in several animal models.A baculovirus pseudotyped with the glycoprotein of vesicular stomatitis virus was used as vector to construct the recombinant baculovirus expressing classical swine fever virus(CSFV) E2 protein under the control of ie1 promoter from white spot syndrome virus.The E2 gene was shown to be efficiently expressed in both insect and mammalian cells.Intramuscular injection of mice with the recombinant baculovirus resulted in the production of high-level CSFV-specific antibodies.Specific lymphoproliferative responses to the CSFV stimulation were induced in the splenocytes of the immunized mice as demonstrated by CFSE staining assay and WST-8 assay.The results indicates that the pseudotyped baculovirus-delivered gene can be a potential non-replicative vaccine against CSFV infection.