Identification of immunodominant fraction of Paramphistomum epiclitum and its evaluation for use in the serodiagnosis of paramphistomosis by ELISA
The present study was undertaken to identify and purify the immunodominant fractions from the excretory secretory (ES) antigen of Paramphistomum epiclitum, a predominant amphistome species infecting ruminants in India. ES antigen was prepared and characterized using SDS-PAGE and Western blot analysi...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Indian Council of Agricultural Research
2018-05-01
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| Series: | Indian Journal of Animal Sciences |
| Subjects: | |
| Online Access: | https://epubs.icar.org.in/index.php/IJAnS/article/view/79827 |
| Summary: | The present study was undertaken to identify and purify the immunodominant fractions from the excretory
secretory (ES) antigen of Paramphistomum epiclitum, a predominant amphistome species infecting ruminants in
India. ES antigen was prepared and characterized using SDS-PAGE and Western blot analysis. Major polypeptides
of 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 ES
antigen of P. epiclitum was initially concentrated using PEG-8000 followed by spin-X UF concentrator with 10
kDa cutoff range and subsequently fractionated by size exclusion chromatography using Sephadex G-25. Cross
reactivity of the P. epiclitum ES antigen was studied with positive sera of F. gigantica and H. contortus. Based on
the cross reactivity profile, the low molecular weight antigenic fraction with 11 kDa polypeptide was selected for
further use in indirect-ELISA. Bovine serum samples (258) were tested with optimized ELISA. Sensitivity of the
ELISA was calculated as 75.0%, while the specificity was 85.0%. The percent positive and negative predictive
values for the test were 70.78 and 87.57%, respectively. |
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| ISSN: | 0367-8318 2394-3327 |