A developed recombinant antigen and monoclonal antibody based solid phase competitive ELISA for detection of foot and mouth disease virus serotype O specific antibodies
- 1ICAR-National Institute on Foot and Mouth Disease, Arugul, Bhubaneswar-752050, Odisha, India
- 2MSc 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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BibTeX
@article{mallick2026developed,
title = {{A developed recombinant antigen and monoclonal antibody based solid phase competitive ELISA for detection of foot and mouth disease virus serotype O specific antibodies}},
author = {S. Mallick and R. P. Singh and J. K. Biswal and J. K. Mohapatra and M. Rout and S. K. Kabi and R. Ranjan},
journal = {Iranian Journal of Veterinary Research},
year = {2026},
month = aug,
doi = {10.22099/ijvr.2026.53916.8090},
url = {https://doi.org/10.22099/ijvr.2026.53916.8090},
note = {CC BY-NC-ND 4.0},
}
RIS
TY - JOUR
TI - A developed recombinant antigen and monoclonal antibody based solid phase competitive ELISA for detection of foot and mouth disease virus serotype O specific antibodies
AU - S. Mallick
AU - R. P. Singh
AU - J. K. Biswal
AU - J. K. Mohapatra
AU - M. Rout
AU - S. K. Kabi
AU - R. Ranjan
PY - 2026
DA - 2026/08/22
JO - Iranian Journal of Veterinary Research
DO - 10.22099/ijvr.2026.53916.8090
UR - https://doi.org/10.22099/ijvr.2026.53916.8090
AB - 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.
SN - 1728-1997
SN - 2252-0589
ER -