Value of Serum and Milk ELISAs in Diagnosis of Fasciolosis in Lactating Buffaloes Using Fasciola gigantica Partially Purified Somatic Antigen

摘要

A developed ELISA was established to diagnose Fasciola gigantica infection in lactating buffaloesby detecting antibodies in both serum and milk samples. An immunogenic fraction was obtained from crudesomatic antigen after fractionation on CNBr-Sepharose 4 B affinity column chromatography and used indetection of fasciolosis in lactating buffaloes. The prevalence of infection in 61 lactating buffaloes wasestablished using serum and milk ELISAs compared with detection of Fasciola eggs in fecal samples. SerumELISA gave the highest diagnostic value (68.9%) followed by milk ELISA (65.6%), while parasitologicalexamination gave 36.1%. By SDS-PAGE, the isolated fraction resolved into five bands of molecular weights 97,84, 65, 21 and 17 KDa. Immunoreactive bands of the isolated fraction were detected by immunoblot usingnaturally infected buffalo sera and positive defatted milk. Two bands of molecular weights 97 and 84 KDa weredetected by positive sera while other two bands of molecular weights 65 and 17 KDa were detected by positivedefatted milk. In conclusion, a partially purified Fasciola gigantica somatic antigen is potent antigen in thediagnosis of fasciolosis in buffaloes using serum and milk ELISAs. Moreover, milk ELISA is suitable as a meanof routine veterinary diagnosis of Fasciola gigantica infection and an alternative to testing sera.