Quantitative anti-PA IgG ELISA : assessment and comparability with the anthrax toxin neutralization assay in goats

dc.contributor.authorNdumnego, Okechukwu Chinazo
dc.contributor.authorCrafford, Jan Ernst
dc.contributor.authorBeyer, Wolfgang
dc.contributor.authorVan Heerden, Henriette
dc.date.accessioned2014-04-17T08:41:38Z
dc.date.available2014-04-17T08:41:38Z
dc.date.issued2013-12-27
dc.description.abstractBACKGROUND: 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.en
dc.description.librarianam2014en
dc.description.librarianab2014
dc.description.sponsorshipThe Deutsche Forschungsgemeinschaft (German Research Foundation)en
dc.description.urihttp://www.biomedcentral.com/1746-6148/9/265en
dc.identifier.citationNdumnego et al.: Quantitative anti-PA IgG ELISA; assessment and comparability with the anthrax toxin neutralization assay in goats. BMC Veterinary Research 2013 9:265.en
dc.identifier.issn1746-6148
dc.identifier.other10.1186/1746-6148-9-265
dc.identifier.urihttp://hdl.handle.net/2263/39669
dc.language.isoenen
dc.publisherBioMed Centralen
dc.relation.requiresAdobe Acrobat Readeren
dc.rights© 2013 Ndumnego et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution Licenseen
dc.subjectProtective antigenen
dc.subjectIndirect ELISAen
dc.subjectToxin neutralization assayen
dc.subjectImmunoglobulinen
dc.subjectSterne vaccineen
dc.subject.lcshGoatsen
dc.subject.lcshAnthraxen
dc.subject.lcshEnzyme-linked immunosorbent assayen
dc.titleQuantitative anti-PA IgG ELISA : assessment and comparability with the anthrax toxin neutralization assay in goatsen
dc.typeArticleen

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