Development and Application of a Double-Antigen Sandwich Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Porcine Circovirus 2

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>A double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.</jats:p>

Evaluation of a Salmonella Enteritidis vaccine and related ELISA for respective induction and assessment of acquired immunity to the vaccine and/or Echinacea purpurea in Awassi Ewes

“…The main disadvantage of using commercial kits is the lack of specificity, since the coating antigens might differ from those of the newly emerging field strains included in the bacterin [19]. The development of an ELISA system for the assessment of vaccine efficiency and/or natural and synthetic immunopotentiators, is a golden standard tool that has been used frequently in the last decades [10,20,21].…”

A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection

“…For the detection of antibodies against PCV2, the systems of antigen production in Escherichia coli (Shang et al 2008, Marcekova et al 2009, Sun et al 2010, Ge et al 2012, insect cells (Nawagitgul et al 2002, Blanchard et al 2003, Liu et al 2004) and insect larvae (Pérez-Martín et al 2008) exhibit more restricted access to techniques when compared with traditional antigen production in cell culture. However, these systems exhibit high purified protein yields (Nawagitgul et al 2002, Spencer et al 2007), which could be used in indirect ELISA methods.…”

Assessing PCV2 antibodies in field pigs vaccinated with different porcine circovirus 2 vaccines using two commercial ELISA systems

“…PCV2 is often associated with a number of disease symptoms in swine including enteritis, respiratory distress, systemic infection, porcine dermatitis, nephritic syndrome, and reproductive problems. These are collectively known as porcine circovirus-associated disease (PCVAD) [ 7 , 14 ]. In contrast, PCV1 has been shown to be nonpathogenic in pigs under experimental conditions [ 2 , 19 ].…”

“…In contrast, PCV1 has been shown to be nonpathogenic in pigs under experimental conditions [ 2 , 19 ]. Since characteristics of PCV2 that affect invasion of the immune system enhance the severity of several diseases in swine [ 7 ], PCV2-associated diseases are generally recognized as having a significant economic impact on the swine industry worldwide [ 8 ]. In addition, PCV2 interacts with other infectious or non-infectious factors ( i.e.…”

“…It is known that ORF2 has greater nucleotide variation than ORF1 [ 11 ]. Therefore, detection of antibodies against PCV2 differs depending on the antigens and detection methods [ 7 , 13 , 14 ].…”