Development of a Double Antigen Tetanus ELISA for Use in Horses
Loading...
Date
2013-05
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
The Ohio State University
Abstract
Horses are commonly vaccinated against tetanus due to the high mortality rate associated with the toxin from the ubiquitous bacterium, Clostridium tetani. The immunogenicity of any vaccine is dependent upon the adjuvant used and the horse’s previous vaccination history. Inconsistent vaccination practices can lead to lack of protection in case of a challenge infection; however, it is hard to quantify the level of protective immunity. Creating an enzyme-linked immunosorbent assay (ELISA) that will accurately detect antibody levels below 0.10 IU/mL will provide researchers with a cost-effective and timely method to precisely measure tetanus-specific antibody titers. A double antigen ELISA method creates a sandwich of the tetanus specific antigens around the sample antibody. The secondary antigen added is labeled with a biotin marker allowing for a colored conjugate to react. This creates a gradation of a colored reaction, allowing for the ELISA plate reader to determine the antibody titers. This is applicable in both commercial and educational settings that are investigating immune responses and vaccine efficacy. The results of this study indicate that the biotin labeled secondary antigen and the tetanus IgG in the serum may not be properly matching, leading to inconsistent IgG readings. One possible explanation for these inconsistent results is that the blocking solution may not have been added in the correct volume, giving inaccurate IgG readings. Suggested improvements to the protocol used in this experiment include plating known amounts of serum to the plate first to then bind the primary and labeled secondary antigen.
Description
Keywords
horses, tetanus, ELISA