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OJVRTM

 

 Online Journal of Veterinary Research

  Volume 11 (2): 1 -11, 2007


Evaluation of ES-F5 serum and milk ELISAs for diagnosis of

Fasciola gigantica infection in lactating buffaloes

 

Ghazy AA, Abdel-Rahman EH, Abd El-Aziz MM

 

 Department of Parasitology and Animal Diseases , Veterinary Research Division, National Research Center, Post Box 12622, 33 El-Tahrir Street, Dokki,, Giza, Cairo, Egypt.


ABSTRACT

 

Ghazy AA, Abdel-Rahman EH, Abd El-Aziz MM., Evaluation of ES-F5 serum and milk ELISAs for diagnosis of Fasciola gigantica infection in lactating buffaloes, Online J Vet Res, 11 (2):1 -11, 2007. Excretory-secretory (ES) products of Fasciola gigantica have been shown to be a source of potential immunodiagnostic antigens. A purification process was adopted to isolate an immunodiagnostic antigen associated with these products. Five fractions were obtained from crude F. gigantica ES products after fractionation by gel filtration chromatography on Sephacryl S-200 column. Fraction 5 (ES-F5) showed higher serodiagnostic potency than other fractions by enzyme linked immunosorbent assay (ELISA) using two-fold serially diluted F. gigantica positive buffaloes sera. The electrophoretic profile of this fraction showed 7 bands of molecular weights 30.0, 26.0, 22.5, 20.0, 17.0, 14.0 and 13.5 KDa as revealed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). For further characterization of the isolated fraction, 6 bands were identified by isoelectric focusing technique (IEF). The isoelectric points (PIs) of the bands were 4.1, 4.9, 5.3, 6.0, 6.3 and 8.3. The immunoreactive bands of ES-F5 antigen were identified by F. gigantica positive buffaloes sera and defatted skim milk using Western immunoblot. Three immunogenic bands (30.0, 14.0 and 13.5 KDa) were identified by both positive sera and milk. A 22.5 KDa band was specifically identified by positive sera, meanwhile a band of 17.0 KDa was exclusively detected by positive milk. A total of 270 liver fluke (F. gigantica) infected and non infected lactating buffaloes, as detected by parasitological examination, were used to evaluate the diagnostic potency of ES-F5 antigen using serum and milk ELISAs. Serum-ELISA gave the highest diagnostic value (62.2%) followed by milk-ELISA (58.2%) compared with the parasitological examination (48.2%). Milk and serum samples from buffaloes, in which infection with F. gigantica was confirmed by detection of eggs in faeces, were used to estimate sensitivity of the ELISAs. Serum and milk ELISAs using ES-F5 antigen proved high sensitivity (99.2 and 97.7%, respectively). The current research introduces five fractions obtained from F. gigantica ES products which were comparatively evaluated for the detection of F. gigantica antibodies. ES-F5 antigen was successfully utilized in the diagnosis of F. gigantica infection in lactating buffaloes, where it recorded high diagnostic sensitivity using serum and milk ELISAs.

 

Keywords: Fasciola gigantica; Lactating buffaloes; ES-F5 antigen; Serum-ELISA; Milk-ELISA.


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