Abstract
Cathepsin-L cysteine proteinase was purified from Fasciola gigantica regurgitant by two-step alcoholic fractionation, followed by ion-exchange chromatography. The purification strategy was evolved to eliminate other contaminating proteins co-precipitating with the purified proteinase during alcoholic fractionation. The enzyme was stable on long-term storage at –20°C rendering it more suitable for field diagnostic use. The purified cathepsin-L cysteine proteinase was assayed for detection of F. gigantica experimental infection in sheep and buffaloes and could detect infection, as early as 4 weeks post-infection by ELISA, Western blotting and Dipstick ELISA. The 28-kDa cathepsin-L cysteine proteinase seems a promising antigen for the diagnosis of tropical fasciolosis in domestic animals.
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Acknowledgement
The authors are highly thankful to the World Bank sponsored National Agricultural Technology Project (MM & CGP) for providing funds to undertake this research. We also thank Director, IVRI and Principal Investigator (NATP-MM) for providing necessary facilities.
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Declaration: Institute animal ethics committee guidelines for use of experimental animals were strictly followed during these experimental studies.
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Yadav, S.C., Saini, M., Raina, O.K. et al. Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants. Parasitol Res 97, 527–534 (2005). https://doi.org/10.1007/s00436-005-1466-8
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DOI: https://doi.org/10.1007/s00436-005-1466-8