Korean Journal of Veterinary Service (한국동물위생학회지)

The Korean Society of Veterinary Service (한국동물위생학회)
권호 표지

Application of PCR for diagnosis of porcine circovirus type 2

돼지 써코바이러스 2형의 진단을 위한 PCR법 적용

  • Park Hyo-Sun (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute) ;
  • Lee Hyo-Sang (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute) ;
  • Na Ki-Bok (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute) ;
  • Lee Kwan-Bok (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute) ;
  • Kang Su-Jung (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute) ;
  • Moon Sun-Hwa (Dangjin branch of the Chungnam Livestock & Veterinary Research Institute)
  • 박효선 (충청남도 축산위생연구소 당진지소) ;
  • 이효상 (충청남도 축산위생연구소 당진지소) ;
  • 나기복 (충청남도 축산위생연구소 당진지소) ;
  • 이관복 (충청남도 축산위생연구소 당진지소) ;
  • 강수정 (충청남도 축산위생연구소 당진지소) ;
  • 문순화 (충청남도 축산위생연구소 당진지소)
  • Published : 2006.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Porcine circovirus (PCV) is a small, nonenveloped virus that contains a single-stranded circular DNA genome of about 1.76 kb and belongs to the family circoviridae. The PCV-2 has been incriminated as the cause of post-weaning multisystemic wasting syndrome (PMWS) , an emerging disease in pigs. In the present study, a PCR assay was applied to detect PCV-2 in tissue samples. The presence of PCV-2 antigen in the porcine tissues was confirmed by indirect immunofluorescence (IIF) with PCV-2 specific monoclonal antibodies. And then DNA extracted from PCV-2 positive tissues was used as a template. One oligonucleotide primer suitable for PCR was selected from a published PCV-2 sequence (Genbank). Amplified PCR product was detected the same fragment lengths of 416 bp as a control. Based on these results, it was suggested that the PCR is a simple and sensitive method for support diagnostic purposes.

Keywords

References

  1. Allan GM, Ells JA. 2000. Porcine circoviruses: A review. J Vet Diagn Invest 12:3-14 https://doi.org/10.1177/104063870001200102
  2. Tischer I, Rasch R, Tochtermann G. 1974. Characterization of papovavirus and picornavirus-like particles in permanent pig kidney cell lines. Zentralbl Bakteriol [Orig A] 226(2):153-167
  3. Cheung AK. 2003. Transcriptional analysis of pocine circovirus. J Virol 305:168-180 https://doi.org/10.1006/viro.2002.1733
  4. Fenaux M, Halbur PG, Gill M, et al. 2000. Genetic characterization of type 2 pocine circovirus(PCV-2) from pigs with post-weaning multisystemic wasting syndrome in different geographic regions of North America and development of a differential PCR-restriction fragment length polymorphism assay to detect and differentiate between infections with PCV-1 and PCV-2. J Clin Microbiol 38:2494-2503
  5. Bassami MR, Berryman D, Wilcox GE, et al. 1998. Psittacine beak and feather disease virus nucleotide sequence analysis and its relationship to porcine circovirus, plant circoviruses, and chicken anaemia virus. Virology 249(2):453-459 https://doi.org/10.1006/viro.1998.9324
  6. Todd D, Niagro FD, Ritchie BW, et al. 1991. Comparison of three animal viruses with circular single-stranded DNA genomes. Arch. Virol 117:129-135 https://doi.org/10.1007/BF01310498
  7. Toischer I, Bode L, Apodaca J, et al. 1995. Presence of antibodies reacting with porcine circovirus in sera of humans, mice, and cattle. Arch Virol 140:1427-1439 https://doi.org/10.1007/BF01322669
  8. Fenaux M, Opriessnig T, Halbur PG, et al. 2003. Immunogenicity and pathogenicity of chimeric infectious DNA clones of pathogenic porcine circovirus type 2(PCV2) and nonpathogenic PCV 1 in weanling pigs. J Virol 77(20):11232-11243 https://doi.org/10.1128/JVI.77.20.11232-11243.2003
  9. Fenaux M, Halbur PG, Haqshenas, et al. 2002. Cloned genomic DNA of type 2 porcine circovirus is infectious when injected directly into the liver and lymph nodes of pigs: Characterization of clinical disease, virus distribution, and pathologic lesions. J Virol 76(2):541-551 https://doi.org/10.1128/JVI.76.2.541-551.2002
  10. Allan GM, McNeilly F, Kennedy S, et al. 1998. Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. J Vet Diagn Invest 10:3-10 https://doi.org/10.1177/104063879801000102
  11. Allan GM, Meehan B, Todd D, et al. 1998. Novel pocine circoviruses from pigs with wasting disease syndromes. Vet Rec 142:467-468
  12. Choi C, Chae C, Clark EG. 2000. Porcine postweaning multisystemic wasting syndrome in Korean pig: Detection of porcine circovirus 2 infection by immunohistochemistry and polymerase chain reaction. J Vet Diagn Invest 12:151-153 https://doi.org/10.1177/104063870001200209
  13. Allan GM, McNeilly F, Meehan BM, et al. 1999. Isolation and characterization of circoviruses from pigs with wasting syndromes in Spain, Denmark and Northern Ireland. Vet Microbiol 66:115-123 https://doi.org/10.1016/S0378-1135(99)00004-8
  14. Hamel AL, Lin LL, Sachvie C, et al. 1998. Nucleotide sequence of porcine circovirus associated with post-weaning multisystemic wasting syndrome in pigs. J Virol 7:5262-5267
  15. Meehan BM, McNeilly F, Todd D, et al. 1998. Characterization of novel circovirus DNAs associated with wasting syndrome in pigs. J Gen Virol 79:2171-2179 https://doi.org/10.1099/0022-1317-79-9-2171
  16. Kim JH, Chung HK, Jung TW, et al. 2002. Postweaning multisystemic wasting syndrome of pigs in Korea: Prevalence, microscopic lesions and coexisting microoganisms. J Vet Med Sci 64(1):57-62 https://doi.org/10.1292/jvms.64.57
  17. McNeilly F, Kennedy S, Moffett D, et al. 1999. A comparison of in situ hybridization and immunohistochemistry for the detection of a new porcine circovirus in formalin-fixed tissues from pigs with post-weaning multisystemic wasting syndrome(PMWS). J Virol Methods 80:2123-2128
  18. Morozov I, Sirinarumitr T, Sorden SD, et al. 1998. Detection of a novel strain of porcine circovirus in pigs with postweaning multisystemic wasting syndrome. J Clin Microbiol 36(9):2535-2541
  19. Krakowka S, Ellis JA, Meehan B, et al. 2000. Viral, wasting syndrome of swine: Experimental reproduction of postweaning multisystemic wasting syndrome in gnotobiotic swine by coinfection with porcine circovirus 2 and porcine parvovirus. Vet Pathol 37:254-263 https://doi.org/10.1354/vp.37-3-254
  20. Mahe D, Blanchard P, Truong C, et al. 2000. Differential recognition of ORF2 protein from type 1 and type 2 porcine circoviruses and identification of immunorelevant epitopes. J Gen Virol 81:1815-1824 https://doi.org/10.1099/0022-1317-81-7-1815
  21. Nawagitgul P, Morozov I, Bolin SR, et al. 2000. Open reading frame 2 of porcine circovirus type 2 encodes a major capsid protein. J Gen Virol 81:2281-2287 https://doi.org/10.1099/0022-1317-81-9-2281
  22. Mager R, Larochelle R, Thibault S, et al. 2000. Experimental transmission of porcine circovirus type 2(PCV2) in weaning pigs : a sequential study. J Comp Pathol 123:258-269 https://doi.org/10.1053/jcpa.2000.0413