Immunodiagnostic potency of homologous antigens for natural Paramphistomum epiclitum infection in small ruminants in plate and paper enzyme linked immunosorbent assay

摘要

The objective of the present work was to standardize and evaluate indirect plate and dot-enzyme linked immunosorbent assay (ELISA) using purified Paramphistomum epiclitum homologous antigens in the small ruminants. Electrophoretic separation of somatic antigen (PeSAg) in reducing condition on 15% polyacrylamide gel resolved into 16 proteins of the molecular weight ranging from 14 - 100 kDa. Two step ethanolic precipitation of supernatant of in-vitro culture of the fluke yielded P. epiclitum excretory-secretory antigen (PeESAg) of molecular weight 28 kDa. The animals (Goats=123; Sheep=91) were broadly kept into post-mortem and faecal examined groups. At many occasion the PeSAg found to cross reacts with other helminths parasites thus minimizing the specificity of the tests and antigens. There was no any direct correlation between the parasites load and ELISA reactivity pattern. The noted prevalence rate after combining the results of postmortem examination and PeESAg based ELISA (plate and paper/dot) was 30.08% (37/123) in goats and 28.57% (26/91) in sheep. While using PeESAg, the calculated overall sensitivity% was 92.86 (goats)/100 (sheep) in both plate and dot-ELISA, specificity% was 91.58 (goats)/91.55 (sheep) in plate ELISA while 88.42 (goats)/92.96 (sheep) in dot-ELISA, positive predictive value% was 76.47 (goats)/76.92 (sheep) in plate ELISA while 70.27 (goats)/80 (sheep) in dot-ELISA and negative predictive value% was 97.75 (goats)/100 (sheep) in plate ELISA while 97.67 (goats)/100 (sheep) in dot-ELISA, these values were optimum for the field sera sample so the tests and PeESAg can be recommended for the detection P. epiclitum infection in the small ruminants.