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Volume 25 Issue 1
February 2010
    Citation: Guo-zhen LIN, Fu-ying ZHENG, Ji-zhang ZHOU, Xiao-an CAO, Xiao-wei GONG, Guang-hua WANG, Chang-qing QIU. An Indirect ELISA of Classical Swine Fever Virus Based on Quadruple Antigenic Epitope Peptide Expressed in E.coli .VIROLOGICA SINICA, 2010, 25(1) : 71-76.  http://dx.doi.org/10.1007/s12250-010-3086-4
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    An Indirect ELISA of Classical Swine Fever Virus Based on Quadruple Antigenic Epitope Peptide Expressed in E.coli

    cstr: 32224.14.s12250-010-3086-4

    • Key Laboratories of Grazing Animal Diseases and Animal Virology, Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China; State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China

    • Corresponding author: Chang-qing QIU, cqqiu@126.com
    • Received Date: 03 July 2009
      Accepted Date: 01 December 2009
      Available online: 01 February 2010
    • 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.
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      2. Jansen van Vuren P, Paweska J T. 2009. Laboratory safe detection of nucleocapsid protein of Rift Valley fever virus in human and animal specimens by a sandwich ELISA. J Virol Methods, 157 (1):15-24.
          doi: 10.1016/j.jviromet.2008.12.003

      3. Lin M, Lin F, Mallory M, et al. 2000. Deletions of structural glycoprotein E2 of classical swine fever virus strain Alfort/187 resolve a linear epitope of monoclonal antibody WH303 and the minimal N-terminal hyperim-mune serum. J Virol, 74: 11619-11625.
          doi: 10.1128/JVI.74.24.11619-11625.2000

      4. Liu S G, Yu X L, Wang C L, et al. 2006. Quadruple antigenic epitope peptide producing immune protection against classical swine fever virus. Vaccine, 24: 7175-7180.
          doi: 10.1016/j.vaccine.2006.06.042

      5. Sambrook J, Fritsch E F, Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd edit, New York: Cold Spring Harbor Laboratory Press, p. 888-897.

      6. Van Gennip H G P, Miedema G K W, Moormann R J M, et al. 2008. Functionality of chimeric E2 glycoproteins of BVDV and CSFV in virus replication. Virology (Auckl), 1: 29-40.

      7. Wu R, Hu S, Xiao, Y, et al. 2007. Development of indirect enzyme-linked immunosorbent assay with nucleoprotein as antigen for detection and quantification of antibodies against avian influenza virus. Vet Res Commun, 31: 631-641.
          doi: 10.1007/s11259-007-3510-x

      8. Yamamoto Y I, Hoshino-Shimizu S, Camargo M E. 1991. A novel IgM-indirect hemagglutination test for the serodiagnosis of acute toxoplasmosis. J Clin Lab Anal, 5: 127-132.
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      9. Yin Z, Liu J H, 1997. Pestivirus, In: Animal Virology, 2 ed, Beijing: Science Press, p 645-664. (in Chinese).

      10. Zhang F, Yu M, Weiland E, et al. 2006. Characterization of epitopes for neutralizing monoclonal antibodies to classical swine fever virus E2 and E (rns) using phage-displayed random peptide library. Arch Virol, 151: 37-54.
          doi: 10.1007/s00705-005-0623-9

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      An Indirect ELISA of Classical Swine Fever Virus Based on Quadruple Antigenic Epitope Peptide Expressed in E.coli

        Corresponding author: Chang-qing QIU, cqqiu@126.com
      • Key Laboratories of Grazing Animal Diseases and Animal Virology, Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China; State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China
      • Received Date: 03 July 2009
      • Accepted Date: 01 December 2009

      Abstract: 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.

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