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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.