• International Journal of Industrial Entomology and Biomaterials
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• 제29권2호
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• pp.207-213
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• 2014

The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

• Asian-Australasian Journal of Animal Sciences
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• 제25권8호
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• pp.1089-1095
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• 2012

Neonatal Fc receptor (FcRn) gene encodes a receptor that binds the Fc region of monomeric immunoglobulin G (IgG) and is responsible for IgG transport and stabilization. In this report, the 8,900 bp porcine FcRn genomic DNA structure was identified and putative FcRn protein included 356 amino acids. Alignment and phylogenetic analysis of the porcine FcRn amino acid sequences with their homologies of other species showed high identity. Tissues expression of FcRn mRNA was detected by real time quantitative polymerase chain reaction (Q-PCR), the results revealed FcRn expressed widely in ten analyzed tissues. One single nucleotide polymorphism (SNP) (HQ026019:g.8526 C>T) in exon6 region of porcine FcRn gene was demonstrated by DNA sequencing analysis. A further analysis of SNP genotypes associated with serum Classical Swine Fever Virus antibody (anti-CSFV) concentration was performed in three pig populations including Large White, Landrace and Songliao Black pig (a Chinese indigenous breed). Our results of statistical analysis showed that the SNP had a highly significant association with the level of anti-CSFV antibody (At d 20; At d 35) in serum (p = 0.008; p = 0.0001). Investigation of expression and polymorphisms of the porcine FcRn gene will help us in further understanding the molecular basis of the antibody regulation pathway in the porcine immune response. All these results indicate that FcRn gene might be regarded as a molecular marker for genetic selection of anti-CSFV antibody level in pig disease resistance breeding programmes.

• Journal of Microbiology and Biotechnology
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• 제25권4호
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• pp.537-546
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• 2015

Classical swine fever (CSF) is a highly contagious disease of pigs caused by CSF virus (CSFV). E2 is the major viral envelope protein of immune dominance that induces neutralizing antibodies and confers protection against CSFV infection. The B/C domains of E2 are variable among CSFV isolates, which could affect immunogenicity and binding to antibodies. We attempted to characterize the epitope recognized by a monoclonal antibody 2B6 (mAb-2B6) raised against the E2 B/C domains of the vaccine C-strain and to examine if mutations in the epitope region would affect antibody binding and viral neutralization. The epitope specific for mAb-2B6 recognition is linear, spanning five residues ⁷⁷⁴DGXNP⁷⁷⁸ in the B/C domains. The residue N777 is indispensable for the specificity. The epitope exists only in group 1 strains, but not in those of group 2. The recombinant viruses containing individual mutations on the epitope region lost the reactivity to mAb-2B6. The mutant virus RecC-N777S had low replication potential, about 10-fold decrease in the yield of progeny virus particles, whereas the mutant virus RecC-P778A reverted to proline upon continuous passaging. The mutations on the mAb-2B6 epitope region did not affect neutralization by anti-C-strain polyclonal sera from pigs. Deletion from aa774 covering the mAb-2B6 epitope, but not that from aa781, also affected binding with the polyclonal antibodies from vaccinated pigs, although the major binding region for the vaccinated antibodies is aa690-773.

• 한국항공우주학회지
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• 제49권1호
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• pp.1-12
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• 2021

본 논문은 복합재 패널에서 압전 작동기를 사용하여 탄성파를 생성하고, 손상에서의 반사된 신호를 압전 감지기에서 탐지하여 손상위치를 추정할 수 있는 알고리즘을 개발하였다. 손상이 없는 신호와 손상이 있는 신호를 비교하여 손상신호를 추정하는 진단적 접근방법을 사용하였다. 신호 상관관계를 이용하여 탄성파의 군속도를 계산하고 압전기 위치정보를 이용하여 손상정보를 추출하였다. 하지만 탄성파의 비선형 특성으로 인해, 손상정보는 다양한 신호의 조합으로 구성되기 때문에, 손상위치를 명확히 구별하기 어렵다. 이에 본 논문에서는 손상에서 반사된 신호정보를 신호 도달거리의 면적으로 변환해서 손상의 중심위치를 찾는 누적함수 특성벡터 알고리즘(CSFV, cumulative summation feature vector)을 새롭게 제안하고, 특성벡터를 손상지수와의 곱으로 표현하는 가시화 기법을 적용하였다. 또한 복합재 패널에서 실험검증을 수행하고, 기존의 알고리즘과의 비교를 통해 제안된 알고리즘이 정확도 높게 손상위치를 검출할 수 있음을 보였다.

• BMB Reports
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• 제40권5호
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• pp.611-616
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• 2007

$E^{rns}$ is an envelope glycoprotein of classical swine fever virus (CSFV) and has an unusual feature of RNase activity. In the present study, we demonstrate that $E^{rns}$ counteracts Newcastle disease virus (NDV)-mediated induction of IFN-$\beta$. For this purpose, $E^{rns}$ fused to the enhanced green fluorescent protein (EGFP) was transiently expressed in porcine kidney 15 (PK15) cells. In luciferase activity assay, $E^{rns}$-EGFP was found to prevent IFN-$\beta$ promoter-driven luciferase expression and block the induction of IFN-$\beta$ promoter mediated by NDV in a dose-dependent manner. Through IFN-specific semi-quantitative RT-PCR detection, obvious decrease of IFN-$\beta$ mRNA in NDV-infected PK15 cells was observed in the presence of $E^{rns}$-EGFP. In contrast, EGFP alone showed none of this block capacity. In addition, $E^{rns}$-EGFP mutations with RNase inactivation were also found to block NDV-mediated induction of IFN-$\beta$. These evidences establish a novel function for CSFV $E^{rns}$ glycoprotein in counteraction of the IFN-$\beta$ induction pathway.

• 대한수의학회지
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• 제43권2호
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• pp.263-270
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• 2003

The objective of the present study was to investigate the use of fluid released from muscle samples as an alternative to serum for ELISA to detect classical swine fever(CSF) virus antibodies in slaughter pigs. The optimal correspondence between serum 1:20 OD values and muscle fluid OD values was achieved at a muscle fluid dilution of 1:2. Significant correlation was found between serum and neck muscle ELISA ($r_s=0.880$, p<0.0001, ${\kappa}=0.82$; specificity of 97.0% and sensitivity 90.6%). The semimembranous muscle showed similar correlation in CSF ELISA($r_s=0.877$, p<0.0001, ${\kappa}=0.75$; specificity of 94.1% and sensitivity 89.1%). High correlation was obtained between serum and mesenteric lymph node in the CSF ELISA ($r_s=0.937$, p<0.0001, ${\kappa}=0.87$; specificity of 97.1% and sensitivity 93.0%). Measmement agreement between serum ELISA and muscle fluid ELISA was calculated and expressed as limits of agreement. The correspondence of ELISA of serum and muscle fluid indicated limits of agreement. Above 95% of all muscle fluid values were distributed within this limits of agreement. Among the samples used for ELISA for detecting CSFV antibodies, mesenteric lymph node had the most correlation and agreement with serum ELISA. F-test for comparison of variances showed no significant difference between the serum and muscle fluid. In conclusion, muscle fluid is a useful postmortem alternative to serum to detect CSFV antibodies.

• 한국동물위생학회지
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• 제43권3호
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• pp.155-159
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• 2020

This study was conducted to investigate the infection of livestock diseases using 500 blood samples from wild boars captured in six cities and one county in northern Gyeonggi province, South Korea. We examined 239 cases of classical swine fever virus (CSFV), and each of 500 cases of foot and mouth disease virus (FMDV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), Mycoplasma hyopneumoniae (MH), Actinobacillus pleuropneumoniae (APP), Pasteurella multocida type A (PMA), Hemophilus parasuis (HP), Salmonella (Sal.) spp. infections. Antibodies were detected against CSFV (23.4%), PRRSV (4.0%), PCV2 (60.4%), MH (3.0%), APP (69.2%), PMA (52.8%), HP (11.8%), and Sal. spp. infections (37.2%). No antibodies were detected against FMDV. As a result of antigenic analysis of 68 positive cases (13.6%) out of 500 PRRS antigen tests, 61 North American cases, 6 European cases, 1 North American-European complex case. PCV2 has 158 positive cases (31.6%) out of 500 antigen tests, and the results indicate that a considerable number of individuals are infected. To our knowledge, this is the first seroprevalence report of MH, APP, PMA, HP, and Sal. spp. infections in wild boars in South Korea.

• 대한수의학회지
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• 제48권1호
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• pp.83-88
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• 2008

In total, 1,142 serum samples were collected from 223 goat flocks rising in five different regions of Korea. These samples were screened for the presence of border disease virus (BDV) antibodies using an enzyme linked immunosorbent assay. Of the 1,142 samples, we found 47 bovine viral diarrhea virus (BVDV) positive cases (4.1%). These positive serum samples were also examined further by using the virus neutralization test against BDV. In addition, samples were tested for both BVDV and classical swine fever virus (CSFV). All of the samples that were seropositive for BDV also demonstrated positive antibody titers against BVDV and CSFV. Due to their common antigenicity, we also determined further the prevalence and carried out virus neutralization test against three pestiviruses: 314 of the goat samples were screened using reverse transcription polymerase chain reaction with primer pairs specific to common pestivirus genome regions. Overall, 1.6% (5/314) of the samples tested was positive for pestivirus. Based on the nucleotide sequence data and the phylogenetic analysis, three isolates were characterized as BVDV type 1 and two isolates as BVDV type 2. However, none of the isolates could be classified as BDV. These results indicate that BVDV-1 and BVDV-2 are the pestivirus strains circulating among Korean goat populations.

• 한국동물위생학회지
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• 제34권2호
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• pp.111-116
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• 2011

Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.

• 한국동물위생학회지
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• 제46권1호
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• pp.59-66
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• 2023

Wild boar is closely related to domestic pigs in terms of genetic homogeneity and the possibility of a source of infection by contact. This study investigated the prevalence of viral diseases from wild boars inhabiting Gyeongsangnam-do, South Korea. A total of 374 blood samples were collected and subjected to antigen tests to detect African swine fever virus (ASFV), Porcine circovirus type-2 (PCV2), Porcine reproductive and respiratory syndrome virus (PRRSV). For seroprevalence, PCV2, PRRS, classical swine fever virus (CSFV), Aujezsky's disease (ADV), and foot and mouth disease virus (FMDV) were investigated. The antigenic analysis revealed 73 positive cases (19.5%) for PCV2, while no positive cases for ASFV and PRRSV. For the antibody test, 225 (60.2%), 2 (0.5%), and 48 (12.8%) cases were detected against PCV2, PRRSV, and CSFV, respectively. There were no antibodies detected against both ADV and FMDV. Our results suggest that the viruses infecting both wild boar and domestic pig, mainly PCV2, are circulating in the wild boar population thus, the consistent monitoring of prevalence in wild boar will be needed for transboundary spillover to the domestic pig.