Skip to main content
PMC home page
Virologica Sinica logoLink to Virologica Sinica
. 2010 Jun 6;25(3):191–198. doi: 10.1007/s12250-010-3085-5

An ELISA based on a truncated soluble ORF2 protein for the detection of PCV2 antibodies in domestic pigs

Shuang-hui Yin 1, Shun-li Yang 1, Hong Tian 1, Jin-yan Wu 1, You-jun Shang 1, Xue-peng Cai 1, Xiang-tao Liu 1,✉
PMCID: PMC8227917  PMID: 20960293

Abstract

Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA) based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.

Key words: Porcine circovirus type 2 (PCV2), Soluble ORF2 protein, ELISA, Antibody detection

Footnotes

Foundation Item: This work was supported by a project from National Key Technology R&D Program in the 11th Five year Plan of China (2006BAD06A12)

References

  • 1.Allan G. M., McNeilly F., Kennedy S., et al. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. J Vet Diagn Invest. 1998;10:3–10. doi: 10.1177/104063879801000102. [DOI] [PubMed] [Google Scholar]
  • 2.Allan G. M., Neilly F. M., Meehan B. M., et al. Isolation and characterisation of circoviruses from pigs with wasting syndromes in Spain, Denmark and Northern Ireland. Vet Microbiol. 1999;66:115–123. doi: 10.1016/S0378-1135(99)00004-8. [DOI] [PubMed] [Google Scholar]
  • 3.Chae C. Postweaning multisystemic wasting syndrome: a review of aetiology diagnosis and pathology. Vet J. 2004;168:41–49. doi: 10.1016/j.tvjl.2003.09.018. [DOI] [PubMed] [Google Scholar]
  • 4.Ellis J., Hassard L., Clark E., et al. Isolation of circovirus from lesions of pigs with postweaning multisystemic wasting syndrome. Can Vet J. 1998;39:44–51. [PMC free article] [PubMed] [Google Scholar]
  • 5.Harding J C. 1997. Post-weaning multisystemic wasting syndrome (PMWS): preliminary epidemiology and clinical presentation. In Proceedings of the 28th Annual Meeting of 197 the American Association of Swine Practitioners. American Association of Swine Practitioners, Quebec City, Canada, 503–507.
  • 6.Hamel A. L., Lin L. L., Nayar G. P. Nucleotide sequence of porcine circovirus associated with postweaning multisystemic wasting syndrome in pigs. J Virol. 1998;72:5262–5267. doi: 10.1128/jvi.72.6.5262-5267.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Krakowka S., Ellis J. A., McNeilly F., et al. Immunologic features of porcine circovirus type 2 infection. Viral Immunol. 2002;15:567–582. doi: 10.1089/088282402320914511. [DOI] [PubMed] [Google Scholar]
  • 8.Liu Q., Tikoo S. K., Babiuk L. A. Nuclear localization of the ORF2 protein encoded by porcine circovirus type 2. Virology. 2001;285:91–99. doi: 10.1006/viro.2001.0922. [DOI] [PubMed] [Google Scholar]
  • 9.Lekcharoensuk P., Morozov I., Paul P. S. Epitope Mapping of the Major Capsid Protein of Type 2 Porcine Circovirus (PCV2) by Using Chimeric PCV1 and PCV2. J Virol. 2004;78(15):8135–8145. doi: 10.1128/JVI.78.15.8135-8145.2004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Liu C. M., Takeshi I. A., Tetsuo N. A., et al. Development of an ELISA based on the bachlovirus-Expressed capsid protein ofn porcine circovirus type 2 as antigen. J Vet Med Sci. 2004;66(3):237–242. doi: 10.1292/jvms.66.237. [DOI] [PubMed] [Google Scholar]
  • 11.Meehan B. M., McNeilly F., Todd D., et al. Characterization of novel circovirus DNAs associated with wasting syndromes in pigs. J Gen Virol. 1998;79:2171–2179. doi: 10.1099/0022-1317-79-9-2171. [DOI] [PubMed] [Google Scholar]
  • 12.Morozov I., Sirinarumitr T., Sorden S. D., et al. Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome. J Clin Microbiol. 1998;36:2535–2541. doi: 10.1128/jcm.36.9.2535-2541.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Magar R., Muller P., Larochelle R. Retrospective serological survey of antibodies to porcine circovirus type 1 and type 2. Can J Vet Res. 2000;64:184–186. [PMC free article] [PubMed] [Google Scholar]
  • 14.Mahe D., Blanchard P., Truong C., et al. Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes. J Gen Virol. 2000;81:1815–1824. doi: 10.1099/0022-1317-81-7-1815. [DOI] [PubMed] [Google Scholar]
  • 15.Nawagitgul P., Morozov I., Bolin S. R., et al. Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein. J Gen Virol. 2000;81(Pt.9):2281–2287. doi: 10.1099/0022-1317-81-9-2281. [DOI] [PubMed] [Google Scholar]
  • 16.Nawagitgul P., Harms P. A., Morozov I., et al. Modified indirect porcine circovirus (PCV) type 2-based and recombinant capsid protein (ORF2)-based enzyme-linked immunosorbent assays for detection of antibodies to PCV. Clin Diagn Lab Immunol. 2002;9:33–40. doi: 10.1128/CDLI.9.1.33-40.2002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Shang S. B., Li Y. F., Guo J. Q., et al. Development and validation of a recombinant capsid protein-based ELISA for detection of antibody to porcine circovirus type 2. Res Vet Sci. 2008;84:150–157. doi: 10.1016/j.rvsc.2007.02.007. [DOI] [PubMed] [Google Scholar]
  • 18.Tischer I., Gelderblom H., Vettermann W., et al. A very small porcine virus with circular single-stranded DNA. Nature. 1982;295:64–66. doi: 10.1038/295064a0. [DOI] [PubMed] [Google Scholar]
  • 19.Tischer I., Mields W., Wolff D., et al. Studies on epidemiology and pathogenicity of porcine circovirus. Vrch Virol. 1986;91:271–276. doi: 10.1007/BF01314286. [DOI] [PubMed] [Google Scholar]
  • 20.Truong C., Mahe D., Blanchard P., et al. Identification of an immunorelevant ORF2 epitope from porcine circovirus type 2 as a serological marker for experimental and natural infection. Arch Virol. 2001;146:1197–1211. doi: 10.1007/s007050170115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Yu X. L., Tu C. C., Li H. W., et al. DNA-mediated protection against classical swine fever virus Vaccine. Vaccine. 2001;19:1520–1525. doi: 10.1016/S0264-410X(00)00334-0. [DOI] [PubMed] [Google Scholar]

Articles from Virologica Sinica are provided here courtesy of Wuhan Institute of Virology, Chinese Academy of Sciences

RESOURCES