Skip to main content

Advertisement

Log in

A Duplex Real-Time PCR Assay for the Simultaneous Detection of Porcine Circovirus 2 and Circovirus 3

  • Research Article
  • Published:
Virologica Sinica

Abstract

Porcine circoviruses (PCV) include PCV1, PCV2, and the new-emerging PCV3. PCV2 is pathogenic to pigs, but the pathogenicity of PCV3 in pigs is debatable. Recently, there have been frequent reports of PCV2 and PCV3 co-infections in clinical samples. Thus, it would be practical to develop a duplex PCR method to detect PCV2 and PCV3 simultaneously. In this study, specific primers and probes were designed to target PCV2 cap and PCV3 rep genes. A duplex real-time PCR method was then developed to detect the two viruses. The assay was found to be highly specific, sensitive, and reproducible for PCV2/3 without cross-reactions with other swine pathogens. The sensitivity of this assay was 2.9 copies for the PCV2 plasmid and 22.5 copies for the PCV3 plasmid. The established assay was then used to detect PCV2/3 infection in 340 clinical samples collected in the first half of 2017. The results showed that the co-infection rate of PCV2/3 in the samples was 27.6%. Our study provides an important tool that can be used to perform urgently needed surveys for the two porcine circoviruses to evaluate their impact on the swine industry.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Fu X, Fang B, Ma J, Liu Y, Bu D, Zhou P, Wang H, Jia K, Zhang G (2018) Insights into the epidemic characteristics and evolutionary history of the novel porcine circovirus type 3 in southern China. Transbound Emerg Dis 65:e296–e303

    Article  CAS  PubMed  Google Scholar 

  • Ku X, Chen F, Li P, Wang Y, Yu X, Fan S, Qian P, Wu M, He Q (2017) Identification and genetic characterization of porcine circovirus type 3 in China. Transbound Emerg Dis 64:703–708

    Article  CAS  PubMed  Google Scholar 

  • Kwon T, Yoo SJ, Park CK, Lyoo YS (2017) Prevalence of novel porcine circovirus 3 in Korean pig populations. Vet Microbiol 207:178–180

    Article  PubMed  Google Scholar 

  • Palinski R, Pineyro P, Shang P, Yuan F, Guo R, Fang Y, Byers E, Hause BM (2017) A novel porcine circovirus distantly related to known circoviruses is associated with porcine dermatitis and nephropathy syndrome and reproductive failure. J Virol 91:e01879-16

    Article  PubMed  Google Scholar 

  • Phan TG, Giannitti F, Rossow S, Marthaler D, Knutson TP, Li L, Deng X, Resende T, Vannucci F, Delwart E (2016) Detection of a novel circovirus PCV3 in pigs with cardiac and multi-systemic inflammation. Virol J 13:184

    Article  PubMed  PubMed Central  Google Scholar 

  • Shen H, Liu X, Zhang P, Wang L, Liu Y, Zhang L, Liang P, Song C (2017) Genome characterization of a porcine circovirus type 3 in South China. Transbound Emerg Dis 65:264–266

    Article  PubMed  Google Scholar 

  • Stadejek T, Wozniak A, Milek D, Biernacka K (2017) First detection of porcine circovirus type 3 on commercial pig farms in Poland. Transbound Emerg Dis 64:1350–1353

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Zhang Y, Liu L, Pang X, Yuan W (2017) Development of a TaqMan-based real-time PCR assay for the specific detection of porcine circovirus 3. J Virol Methods 248:177–180

    Article  CAS  PubMed  Google Scholar 

  • Yun JJ, Heisler LE, Hwang II, Wilkins O, Lau SK, Hyrcza M, Jayabalasingham B, Jin J, McLaurin J, Tsao MS, Der SD (2006) Genomic DNA functions as a universal external standard in quantitative real-time PCR. Nucleic Acids Res 34:e85

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhai SL, Chen SN, Xu ZH, Tang MH, Wang FG, Li XJ, Sun BB, Deng SF, Hu J, Lv DH, Wen XH, Yuan J, Luo ML, Wei WK (2014) Porcine circovirus type 2 in China: an update on and insights to its prevalence and control. Virol J 11:88

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by Grants from the National Key Research and Development Program (2016YFD0500703), Major Science and Technology Projects in Henan Province (171100110200), and Luoyang HeLuo Talent Plan (Dr. Kegong Tian).

Author information

Authors and Affiliations

Authors

Contributions

MS and KGT designed the study. XDL and MMQ performed the experiments and wrote the manuscript.

Corresponding authors

Correspondence to Ming Sun or Kegong Tian.

Ethics declarations

Conflict of interest

The authors declare that they do not have any conflict of interest.

Animal and Human Rights Statement

All animal trials were performed in strict accordance with the regulations in the guide for the care and use of laboratory animals of the National Research Center for Veterinary Medicine.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 307 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Li, X., Qiao, M., Sun, M. et al. A Duplex Real-Time PCR Assay for the Simultaneous Detection of Porcine Circovirus 2 and Circovirus 3. Virol. Sin. 33, 181–186 (2018). https://doi.org/10.1007/s12250-018-0025-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12250-018-0025-2

Keywords

Navigation