DEVELOPMENT OF SERODIAGNOSTIC ASSAYS FOR DETECTION OF SCHISTOSOMA SPINDALE IN CATTLE
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Date
2021
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Abstract
Serodiagnostic assays such as Enzyme linked immunosorbent assay
(ELISA) and Flow through technique (FTT) were standardized and evaluated by
using excretory-secretory (ES) antigen for detection of Schistosoma spindale
infection in cattle. ES antigen of S. spindale was prepared by incubating adult
worms in IX PBS overnight. The ES antigen prepared was concentrated and the
protein content was estimated by Bradford assay.
Protein profiling of 5. spindale ES antigen was carried out by SDS-PAGE
which revealed four polypeptide bands of molecular sizes 14, 28, 57 and 66 kDa.
Immunoreactive bands of molecular weight size 28, 57 and 66 kDa were
confirmed by probing ES antigen with hyper immune sera of rabbits in Western
blotting.
Serodiagnostic assays such as ELISA and FTT were standardized for the
detection of visceral schistosomosis. A total of 180 mesentery and serum samples
were collected from cattle and 57 were found positive for S. spindale infection.
Out of the 57 positive samples, 54 and 53 were sero-positive by ELISA
and FTT respectively. Kappa statistics showed existence of perfect agreement and
Chi-Square test disclosed no significant difference between ELISA and FTT when
compared with the gold standard mesentery examination.
The sensitivity, specificity, positive predictive value, negative predictive
value, accuracy, concordance, false positive rate, false negative rate, kappa value
and Youden index of ELISA in detecting serum antibodies in cattle was 94.73 per
cent, 95.12 per cent, 90 per cent, 97.5 per cent, 95 per cent, 94.48 per cent, 4.88
per cent, 5.27 per cent, 0.8743 and 0.89 respectively. The sensitivity, specificity,
positive predictive value, negative predictive value, accuracy, concordance, false
positive rate, false negative rate, kappa value and Youden index of FTT in
detecting serum antibodies in cattle was 92.98 per cent, 96.74 per cent, 92.98 per
cent, 96.74 per cent, 95.55 per cent, 95.56 per cent, 3.26 per cent, 7.02 per cent,
0.8973 and 0.892 respectively.
Flow through technique, which was not used in diagnosis of visceral
schistosomosis found to be the more accurate, specific, simplest and rapid with
less expertise and easy applicability in field conditions and hence an ideal pen side
diagnostic kit.