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Detection of Anti-PCV2 Antibodies


Detection of PCV2 Nucleic Acids

Detection of PCV2 Virus or Viral Antigen

Further Characterization of PCV2 Isolate

Available Tests


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Serum-virus neutralization (SVN) assay
The basis of the SVN assay is that it detects the presence of antibodies that have the ability to prevent virus from attaching to and/or infecting cells. Neutralizing antibody assays for PCV2 have been described in the literature (Meerts et al., 2006, 2005c; Pogranichnyy et al., 2000). Neutralizing antibodies were detected between 15 (Meerts et al., 2005c) and 28(Pogranichnyy et al., 2000) days post PCV2-infection and were correlated with protection or clearance of PCV2 infection in gnotobiotic pigs (Meerts et al., 2005c).

Indirect immunoperoxidase monolayer assay (IPMA)

The IPMA is similar to the IFA with the exception that the antibody used is a peroxidase-conjugated anti-swine IgG. IPMA for PCV2 is widely used (Ellis et al., 1998; Balasch et al., 1999). An interlaboratory testing comparing IFA and IPMA results on the same 20 serum samples performed in different laboratories in Europe and Canada found a wide variation in detected titers between laboratories (McNair et al., 2004). In general, IPMA gave higher titers than IFA, and paraformaldehyde used as fixative gave higher titers than did acetone or ethyl alcohol (McNair et al., 2004).

Indirect immunofluorescence (IFA) assay

This assay detects the ability of the antibody in the serum to bind to a fixed monolayer of virus-infected cells. The specific antibodies are labeled with fluorescein-conjugated anti-swine-IgG. IFA assays for PCV2 have been described in the literature (Allan et al., 1998; Pogranichnyy et al., 2000; Tischer et al, 1995). Recently, an IFA assay based on an ORF2 protein has been described(Racine et al., 2004) and it was found that the regular whole PCV2-based IFA had only a 57.1% relative sensitivity compared to the ORF-2 protein based IFA. IFAs have also been described for antibodies against non-pathogenic PCV1 (Fenaux et al., 2003) and appear to be specific. Studies have shown that there is a low level of cross-reactivity between PCV1 and PCV2 on IFA test (Allan et al., 1998, Pogranichnyy et al., 2000).

Enzyme-linked immunosorbent assay (ELISA)


There are several peer-reviewed articles describing PCV2 ELISA assays (Blanchard et al., 2003; Liu et al., 2004; Nawagitgul et al., 2002). Recently, commercially available IgG and IgM PCV2-ELISA assays have been introduced in Europe (Ingezim PCV IgG® and Ingezim PCV IgM®, Ingenasa, Madrid, Spain). Combined IgG and IgM values might be useful to determine the timing of PCV2-infection: IgM values ≥ IgG values: early active infection (within the first 21 DPI); IgM values < IgG values: active infection (approximately between 20 to 50 DPI); High IgG values and negative IgM values: late or resolving infection of convalescent (approximately 2 months after infection) (Segalés et al., 2005). A variant of the regular ELISA assay is the competitive (blocking) ELISA (Walker et al., 2000). A competitive ELISA specific for PCV2-antibodies is available in Europe (Synbiotics, Europe, Lyon, France). An antibody-detection blocking ELISA to detect PCV2-specific antibodies on feces is also described(Lopez et al., 2005) and is commercially available in Europe (Synbiotics Europe, Lyon, France).


Allan GM, McNeilly F, Kennedy S, Daft B, Clarke EG, Ellis JA, Haines DM, Meehan BM, Adair BM: Isolation of porcine circovirus-like viruses from pigs with a wasting disease in the USA and Europe. J Vet Diagn Invest. 10:3-10, 1998

Balasch M, Segalés J, Rosell C, Domingo M, Mankertz A, Urniza A, Plana-Durán J: Experimental inoculation of conventional pigs with tissue homogenates from pigs with post-weaning multisystemic wasting syndrome. J Comp Pathol. 121:139-148, 1999

Blanchard P, Mahé D, Cariolet R, Truong C, Le Dimna M, Arnauld C, Rose N, Eveno E, Albina E, Madec F, Jestin A: An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome. Vet Microbiol. 94:183-194, 2003

Fenaux M, Opriessnig T, Halbur PG, Meng XJ: Immunogenicity and pathogenicity of chimeric infectious DNA clones of pathogenic porcine circovirus type 2 (PCV2) and nonpathogenic PCV1 in weanling pigs. J Virol. 77:11232-11243, 2003

Liu C, Ihara T, Nunoya T, Ueda S: Development of an ELISA based on the baculovirus-expressed capsid protein of porcine circovirus type 2 as antigen. J Vet Med Sci. 66:237-242, 2004

Lopez P, Guillossou S, Deshaies E, Brajon N, Bestul K, Leterme S: Porcine circovirus type 2: enzyme linked immunosorbent assay (ELISA) for the detection of antigen and antibodies in feces. In: Proc Intern Conf “Animal Circoviruses and Associated Diseases”, Belfast, UK, p 91, 2005

McNair I, Marshall M, McNeilly F, Bøtner A, Ladekjær-Mikkelsen AS, Vincent I, Herrmann B, Sanchez R, Rhodes C: Interlaboratory testing of porcine sera for antibodies to porcine circovirus type 2. J Vet Diagn Invest. 16:164-166, 2004

Meerts P, Misinzo G, Lefebvre D, Nielsen J, Bøtner A, Kristensen CS, Nauwynck HJ: Correlation between the presence of neutralizing antibodies against porcine circovirus type 2 (PCV2) and protection against replication of the virus and development of PCV2-assoicated disease. BMC Vet Res. 30;2:6, 2006

Meerts P, Van Gucht S, Cox E, Vandebosch A, Nauwynck HJ:Correlation between type of adaptive immune response against porcine circovirus type 2 and level of virus replication. Viral Immunol. 18:333-341, 2005c

Nawagitgul P, Harms PA, Morozov I, Thacker BJ, Sorden SD, Lekcharoensuk C, Paul PS: 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. 9:33-40, 2002

Pogranichnyy RM, Yoon KJ, Harms PA, Swenson SL, Zimmerman JJ, Sorden SD: Characterization of immune response of young pigs to porcine circovirus type 2 infection. Viral Immunol. 13:143-153, 2000

Racine S, Kheyar A, Gagnon CA, Charbonneau B, Dea S:Eucaryotic expression of the nucleocapsid protein gene of porcine circovirus type 2 and use of the protein in an indirect immunofluorescence assay for serological diagnosis of postweaning multisystemic wasting syndrome in pigs. Clin Diagn Lab Immunol. 11:736-741, 2004

Segalés J, Rodríguez J, Resendes A, Balasch M, Sanz AJ, Plana-Durán J, Venteo A: Humoral immune responses and correlation with viraemia in pigs subclinically infected with porcine circovirus type 2. In: Proc Intern Conf “Animal Circoviruses and Associated Diseases”, Belfast, UK, p 61, 2005

Tischer I, Bode L, Apodaca J, Timm H, Peters D, Rasch R, Pociuli S, Gerike E:Presence of antibodies reacting with porcine circovirus in sera of humans, mice, and cattle. Arch Virol. 140:1427-1439, 1995

Walker IW, Konoby CA, Jewhurst VA, McNair I, McNeilly F, Meehan BM, Cottrell TS, Ellis JA, Allan GM: Development and application of a competitive enzyme-linked immunosorbent assay for the detection of serum antibodies to porcine circovirus type 2. J Vet Diagn Invest. 12:400-405, 2000