Abstract:
BACKGROUND: Presently, few data exist on the level and duration of anti-protective antigen (PA) IgG in vaccinated
livestock. Various adaptation of enzyme-linked immunosorbent assays (ELISAs) have been developed in studies
to assess immune response following vaccination, albeit mostly in laboratory rodent models. The quantitative
anti-anthrax IgG ELISA in this study describes a method of enumerating the concentration of anti-PA specific IgG
present in sera of immunized goats, with the aid of an affinity-purified caprine polyclonal anti-anthrax PA-83 IgG
standard. This was compared with the anthrax toxin neutralization assay (TNA) which measures a functional subset of
toxin neutralizing anti-PA IgG.
RESULTS: The measured concentrations obtained in the standard curve correlated with the known concentration at
each dilution. Percentage recovery of the standard concentrations ranged from 89 to 98% (lower and upper asymptote
respectively). Mean correlation coefficient (r2) of the standard curve was 0.998. Evaluation of the intra-assay coefficient
of variation showed ranges of 0.23-16.90% and 0.40-12.46% for days 28 and 140 sera samples respectively, following
vaccination. The mean inter-assay coefficient of variation for triplicate samples repeated on 5 different days was 18.53
and 12.17% for days 28 and 140 sera samples respectively. Spearman’s rank correlation of log-transformed IgG
concentrations and TNA titres showed strong positive correlation (rs = 0.942; p = 0.01).
CONCLUSION: This study provides evidence that an indirect ELISA can be used for the quantification of anti-anthrax PA
IgG in goats with the added advantage of using single dilutions to save time and resources. The use of such related
immunoassays can serve as potential adjuncts to potency tests for Sterne and other vaccine types under development
in ruminant species. This is the first report on the correlation of polyclonal anti-anthrax PA83 antibody with the TNA in
goats.
