A developed recombinant antigen and monoclonal antibody based solid phase competitive ELISA for detection of foot and mouth disease virus serotype O specific antibodies

Document Type : Short paper

Authors

1 ICAR-National Institute on Foot and Mouth Disease, Arugul, Bhubaneswar-752050, Odisha, India

2 MSc in Biotechnology, ICAR-National Institute on Foot and Mouth Disease, Arugul, Bhubaneswar-752050, Odisha, India

Abstract

Background: Foot and mouth disease (FMD) is ahighly contagious and economically burdensome viral disease of cloven-hoofed animals. Out of the seven FMD virus (FMDV) serotypes, type O is one of the most prevalent serotype in several parts of the world. Aims: This study aimed to develop a recombinant capsid polyprotein (rP1) and monoclonal antibody (MAb)-based solid phase competitive ELISA (rP1-MAb-SPCE) to detect anti-FMDV/O antibodies in serum samples. Methods: For the development of rP1-MAb-SPCE the concentrations of rP1, MAb and dilution of serum were optimized. Percent inhibition (PI) cut-off was determined based on ROC curve analysis of cattle sera (n=53) of known FMD vaccination status with reference to the gold standard virus neutralization test (VNT). The assay performance was further validated in comparison to the conventional polyclonal antibody (PAb)-based SPCE using FMDV-positive and negative serum samples (n=612) obtained from multiple species (cattle, goat, pig, fallow deer, antelope, giraffe and African buffalo). Results: At the selected cut-off value of 30.77 PI, the rP1-MAb-SPCE exhibited 86.21% diagnostic sensitivity and 95.83% diagnostic specificity. A significant correlation (r=0.84; P<0.0001) was observed between the PI values of sera tested by rP1-MAb-SPCE and PAb-based SPCE and also the limits of detection of both of the assays were comparable. Repeatability tests demonstrated that the coefficients of variation within and between runs were <10% and <14%, respectively. Conclusion: The developed rP1-MAb-SPCE has the potential to facilitate the detection of antibodies against FMDV serotype O while mitigating biosecurity concerns. However, further validation involving a larger serum cohort is essential to confirm the assay’s robustness.

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References

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