• Title/Summary/Keyword: A classical swine fever vaccine virus

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Classical Swine Fever Virus: Discrimination Between Vaccine Strains and Korean Field Viruses by Real-time RT-PCR

  • Park, Suk-jun;Cho, Ho-seong;A.W.E. Effendy;Kim, Yong-hwan;Park, Nam-yong
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.34-34
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    • 2003
  • Classical swine fever (CSF) is a contagious disease of swine with serious economic losses in pig industry [1]. The disease is caused by CSFV which belongs to the viruses of bovine viral diarrhea (BVDV) and border disease virus (BDV) make up the Pestivirus genus within the family Flaviviridae [2]. Attenuated Korean LOM strains were used in Korea. For these reasons a practical approach for discrimination between vaccine and field strains is needed. Here, we described the deveopment of real-time RT-PCR to discriminate between vaccine strains and Korean field viruses of CSFV. (omitted)

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Sequence Analysis of the Gene Encoding gp55 Protein of Suri Strain, an Attenuated Classical Swine Fever (Hog Cholera) Virus (Classical Swine Fever (Hog Cholera) Virus 약독순화주 (Suri 주)의 gp55 Gnen 염기서열 분석)

  • Kim, Kui-Hyun;Chang, Kyung-Soo;Kang, Kyong-Im;Lee, Byung-Hyung;Park, Jong-Hyeon;An, Soo-Hwan;Jun, Moo-Hyung
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.303-316
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    • 1998
  • An attenuated classical swine fever virus (CSFV), Suri strain, is a variant derived from a vaccine virus, LOM strain. This study was performed to elucidate the molecular biologcal properties of CSFV Suri strain, and to obtain the basic data for molecular epidemiological approaches for the disease. The truncated form of gp55 gene without the C-terminal transmembrane domain, in size of 1,023bp, was amplified by RT-PCR and sequenced by dye terminator cyclic sequencing method, and inserted into BamHI site of pAcGP67B baculovirus vector, establishing a cloned pAcHEG plasmid. By the nucleotide sequences determined, 341 amino acid sequences were predicted. As compared the nucleotide and amino acid sequences of gp55 of Suri with the various CSFV, Suri strain showed the high homology over 99.1% with ALD and LOM strains, but comparably the lower homology with Alfort and Brescia. In comparison of amino acid sequence in variable domain of gp55 protein, the similar tendency of homology was observed. In hydrophobicity analysis, all of four CSFV strains revealed the analogous patterns of hydrophobicity. The numbers and locations of N-glycosylation site and cysteine residues in gp55 were analyzed, those of Suri strain being coincident with ALD and LOM strains. The results suggest that gp55 in Suri strain has the high similarity to those in ALD and LOM strains in terms of the nucleotide and amino acid sequences and the functional properties of gp55 protein.

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Continuous Passaging of a Recombinant C-Strain Virus in PK-15 Cells Selects Culture-Adapted Variants that Showed Enhanced Replication but Failed to Induce Fever in Rabbits

  • Tong, Chao;Chen, Ning;Liao, Xun;Yuan, Xuemei;Sun, Mengjiao;Li, Xiaoliang;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.27 no.9
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    • pp.1701-1710
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    • 2017
  • Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

Epidemiological characteristics of 2002 outbreak of classical swine fever in Korea (2002년 한국에서 발생한 돼지콜레라의 역학적 특성)

  • Park, Choi-Kyu;Song, Jae-Young;Wee, Sung-Hwan;Lee, Eune-Sub;Yoon, Hachung;Moon, Oun-Kyeong;Choi, Eun-Jin;Nam, Hyang-Mi
    • Korean Journal of Veterinary Research
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    • v.46 no.2
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    • pp.107-117
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    • 2006
  • This paper described the epidemiological characteristics of 2002 outbreak of classical swine fever (CSF) in Korea. A total of thirteen CSF-infected farms could be classified into two clusters according to the location and time of outbreak. Two farms located in the same county of Gangwon province and 11 farms located in several different districts of Incheon metropolitan/Gyeonggi province were identified as CSF-infected from April 16 to 30 and from October 7 to December 21 in 2002, respectively. As the result of epidemiological analysis, the two clusters of outbreaks were turned out to be independent epidemics which had different sources of virus introduction. Three farms were found to have been infected primarily; one located in Cheolwon county of Gangwon province and two located in Kangwha county of Incheon metropolitan area. The most likely factors of virus introduction into these primary infected farms were considered to be direct or indirect contact by foreign workers and/or owners of the infected farms who had come back from traveling in China before outbreaks. This was supported by the genetic typing of CSF viruses isolated from the pigs of infected farms. All the virus isolates of 2002 outbreak were found to be genetic type 2, whereas the viruses isolated before 2000 were type 3 and the reference strains, such as attenuated live vaccine virus (LOM strain) and high virulent challenge virus (ALD strain), were type 1. Accordingly, we concluded that the 2002 CSF outbreak must have been caused by a newly introduced virus from overseas and the type 3 virus must have been eradicated after the last outbreak of 1999 by the national CSF eradication campaign which were implemented since 1996. Based on the combined analysis of epidemiological data and genetic typing, the transmission routes of classical swine fever virus were found to be the movement of vehicles (60%) and persons (10%), neighbourhood spread (20%) and unknown (10%). It is expected that the analyzed data and findings of classical swine fever outbreak epidemic could be very useful to establish the disease control and eradication program for the country in the future.

The Epitope Recognized by Monoclonal Antibody 2B6 in the B/C Domains of Classical Swine Fever Virus Glycoprotein E2 Affects Viral Binding to Hyperimmune Sera and Replication

  • Tong, Chao;Chen, Ning;Liao, Xun;Xie, Wenqi;Li, Dejiang;Li, Xiaoliang;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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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.

Epidemiological surveys of an outbreak of false positive classical swine fever in Gyeongbuk province (의사 돼지콜레라 발생농장 역학조사)

  • Park, No-Chan;Cho, Kwang-Hyun;Kim, Young-Hoan;Kim, Soon-Tae;Kim, Sung-Kuk;Park, In-Hwa;Cho, Min-Hee;Oh, Gang-Hee;Son, Jae-Kweon;Jyeong, Jong-Sik
    • Korean Journal of Veterinary Service
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    • v.26 no.2
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    • pp.163-184
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    • 2003
  • This study was conducted to survey the farm which suffered from disease similar to classical swine fever(CSF) in Gyeongbuk province. Clinical signs appeared first in a few number of growing pigs which showed specific signs of diarrhea, depression, sleepiness, and reluctance to get up or to eat. Younger piglets may have appeared chilled, shiver and huddle together. As the disease progresses the affected pig's skin went red and purple. In histopathological signs, there were many haemorrhages throughout the body and larger haemorrhages in some organs such as lymph nodes. And there is a precipitous fall in the number of circulating leukocytes in the blood. In spite of insisting of farmer which did not vaccinate to classical swine fever, significant antibody production was detected in these affected pigs at enzyme-linked immuonsorbent assay. According to the above results at first glance, these affected pig suspected with CSF in clinical signs and histopathological lesions only. Because the symptoms and post-mortem picture were very similar to CSF, these false positive results would have been dangerous to diagnostician. But by reverse transcriptase polymerase chain reaction(RT-PCR) and comparative nucleotide sequence analysis, the disease was correctly diagnosed with post-weaning multisystemic wasting syndrome(PMWS) and porcine reproductive and respiratory syndrome(PRRS) compoundly. And the antigen which were detected the lesion similar to CSF virus, was confirmed with LOM vaccine strain of CSF. In most national CSF eradication program and in countries which are free of the CSF virus, vaccination against CSF is not practiced and generally is not allowed. But now in Korea, routine vaccination is practiced because of outbreaking the CSF repeatedly. When CSF is diagnosed the whole herd and other in contact animal are slaughtered continuously.

Mucosal Immune Response and Adjuvant Activity of Genetically Fused Escherichia coli Heat-Labile Toxin B Subunit

  • Lee, Yung-Gi;Kang, Hyung-Sik;Lee, Cheong-Ho;Paik, Sang-Gi
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.490-497
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    • 2004
  • Although the E. coli heat-labile enterotoxin B subunit (LTB) is known to be a potent mucosal adjuvant towards co-administrated unrelated antigens and immunoregulator in T-helper 1-type-mediated autoimmune diseases, a more efficient and useful LTB is still required for prospective vaccine adjuvants. To determine whether a novel chimeric LTB subunit would produce an enhanced mucosal adjuvant activity and immune response, a number of LTB subunits were genetically fused with chimeric proteins using the epitope genes of the envelope glycoprotein E2 (gp51-54) from the classical swine fever virus (CSFV). It was found that the total serum immunoglobulin (Ig) levels of BALB/c mice orally immunized with chimeric proteins containing an N-terminal linked LTB subunit (LE1, LE2, and LE3) were higher than those of mice immunized with LTB, E2 epitope, and chimeric proteins that contained a C-terminal linked LTB subunit. In particular, immunization with LE1 markedly increased both the total serum Ig and fecal IgA level compared to immunization with LTB or the E2 epitope. Accordingly, the current results demonstrated that the LTB subunit in a chimeric protein exhibited a strong mucosal adjuvant effect as a carrier molecule, while the chimeric protein containing the LTB subunit stimulated the mucosal immune system by mediating the induction of antigen-specific serum Ig and mucosal IgA. Consequently, an LE1-mediated mucosal response may contribute to the development of effective antidiarrhea vaccine adjuvants.