Abstract
The present study was undertaken to identify and purify the immunodominant fractions from the excretorysecretory (ES) antigen of Paramphistomum epiclitum, a predominant amphistome species infecting ruminants inIndia. ES antigen was prepared and characterized using SDS-PAGE and Western blot analysis. Major polypeptidesof molecular weight 11, 22, 28, 31, 33, 39, 52, 59, 63 and 72 kDa were visualized in SDS-PAGE. Polypeptides (9)of 11, 14, 16, 22, 31, 33, 39, 63 and 72 kDa showed immunoreactivity in Western blot analysis. The whole ESantigen of P. epiclitum was initially concentrated using PEG-8000 followed by spin-X UF concentrator with 10kDa cutoff range and subsequently fractionated by size exclusion chromatography using Sephadex G-25. Crossreactivity of the P. epiclitum ES antigen was studied with positive sera of F. gigantica and H. contortus. Based onthe cross reactivity profile, the low molecular weight antigenic fraction with 11 kDa polypeptide was selected forfurther use in indirect-ELISA. Bovine serum samples (258) were tested with optimized ELISA. Sensitivity of theELISA was calculated as 75.0%, while the specificity was 85.0%. The percent positive and negative predictivevalues for the test were 70.78 and 87.57%, respectively.
Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
