DEVELOPMENT OF INDIRECT PCV2B-BASED ELISA FOR DETECTION PCV2 ANTIBODIES

Abstract

This trial was a development of an indirect enzyme-linked immunosorbent assay (ELISA) using whole particle of porcine circovirus type 2 (PCV2) subtype B as antigen and to validate these diagnostic performances. PCV2b antigen was prepared by propagation in SST-SW cell and harvested from cell cultured followed by purifying with sucrose gradient technique. The ELISA was optimized by using checker board titration. As a result, the cut-off value was set at 0.39 (450 nm) with 92% and 100% sensitivity and specificity, respectively (n = 100). Median value of %CV of intra-plate at 3.62 and 6.21 and inter-plate at 7.16 and 6.34 for negative (n = 10) and positive sample (n = 10), respectively. The observed agreement was 89.34% with 0.757 kappa value (n = 657) when compared to IPMA and correlation of ELISA and immunperoxidase monolayer assay (IPMA) resulted in Spearman correlation of coefficient (r) at 0.74 (p < 0.001) (n=20). Based on validation results, indirect PCV2b-based ELISA could may be used for detection of antibodies against PCV2 infection as alternative to IPMA.