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

BMC Veterinary Research

Henriette van Heerden¹  Wolfgang Beyer²  Jannie Crafford¹  Okechukwu C Ndumnego¹ 

[1] Department of Veterinary Tropical Diseases, University of Pretoria, Onderstepoort 0110, South Africa;Institute of Environmental and Animal Hygiene, University of Hohenheim, Stuttgart, Germany

关键词: Goats;    Sterne vaccine;    Immunoglobulin;    Anthrax;    Toxin neutralization assay;    Indirect ELISA;    Protective antigen;   

Others  :  1119373
DOI  :  10.1186/1746-6148-9-265

 received in 2013-09-23, accepted in 2013-12-20,  发布年份 2013

【 摘 要 】

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 (r_s = 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.

【 授权许可】

   
2013 Ndumnego et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

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