Development of a double sandwich fluorescent ELISA to detect rattlesnake venom in biological samples from horses with a clinical diagnosis of rattlesnake bite.

The study aims to establish a test to detect rattlesnake venom in horses, which could help decide the necessity of expensive and potentially harmful anti-venom treatment. To this end, it developed a double sandwich fluorescent ELISA (enzyme-linked immunosorbent assay) and found its effectiveness in detecting venom in both urine and bite site samples.

Study Objective and Method

• The main objective of this research was to develop an ELISA, a test that measures the antibodies in the blood, to detect rattlesnake venom in biological samples from horses clinically diagnosed with rattlesnake bites. Since treatment of horses with anti-venom can be quite expensive and risk potential negative side effects, a quantifiable test is beneficial to determine if such a treatment is necessary.
• A double sandwich fluorescent ELISA was developed for this purpose. The methodology involves the application of two layers of antibodies to sandwich the antigen (in this case, the rattlesnake venom), with the aid of a fluorescent dye for enhanced detection.

Participants and Sample Collection

• For the study, nineteen horses, clinically diagnosed with rattlesnake bites, were enrolled. Of these, urine samples were available from all 19 while bite site samples were obtainable from only nine.

Findings and Future Work

• Post the assay’s implementation, it successfully detected venom in five of the nine bite site samples and twelve of the nineteen urine samples. These results suggest that the assay has the ability to successfully detect rattlesnake venom in horses.
• Future work needs to establish a correlation between venom concentration and clinical outcome to decipher its usefulness as a treatment guide. This would require the determination of the first peak venom concentration.
• To define a venom elimination pattern in horses and to ascertain the best sample collection time for estimating the maximum venom dose, a more frequent and consistent sample collection is necessitated.

Conclusion

• The research report illustrates the successful development of a double sandwich fluorescent ELISA that can detect the presence of rattlesnake venom in horses’ urine and at the bite site. If further refined and validated, this development has the potential to provide a high impact on the treatment decision-making for rattlesnake bites in horses.