title: Development and application of a double-antigen sandwich enzyme-linked immunosorbent assay for detection of antibodies to porcine circovirus 2.
abstract: 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.
authors: College of Veterinary Medicine, Hunan Agricultural University, Changsha, China.
authors: Meng
authors: M
authors: Ge
authors: Wei
authors: W
authors: Luo
authors: Daliang
authors: D
authors: Jiang
authors: Runcheng
authors: R
authors: Li
authors: Wenwei
authors: W
authors: Zhao
authors: Guoliang
authors: G
authors: Chen
authors: Xingdong
authors: X
authors: Yang
authors: Xinglong
authors: X
authors: Yu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D036022:Capsid Proteins / Q000235:genetics; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D017925:Circovirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D036022:Capsid Proteins; D011994:Recombinant Proteins