Fasciola gigantica cathepsin-L cysteine proteinase in the detection of early experimental fasciolosis in ruminants.
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 degrees 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.
Yadav
S C
SC
Division of Parasitology, IVRI, Izatnagar, India.
Saini
M
M
Raina
O K
OK
Nambi
P A
PA
Jadav
K
K
Sriveny
D
D
D000438:Alcohols; D000818:Animals; D000833:Animals, Suckling; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015415:Biomarkers / Q000097:blood; D015153:Blotting, Western / Q000379:methods; D002020:Buffaloes; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005209:Fasciola / Q000276:immunology; D005211:Fascioliasis / Q000175:diagnosis*; D005260:Female; D008297:Male; D008970:Molecular Weight; D012698:Serologic Tests; D012756:Sheep
D000438:Alcohols; D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015415:Biomarkers; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L