Abstract
Porcine Circovirus 2 (PCV2) is the major causative agent of postweaning multisystemic wasting syndrome (PMWS) of swine and is one of the reasons for severe economic loss in swine industry. In India, there is a considerable prevalence rate of PCV2 infection in pig population, PCV2d being the most prominent genotype. Proper sero-diagnosis and sero-surveillance of the disease is formulated as an effective control measure. In this study, a recombinant capsid protein-based single serum dilution indirect ELISA was developed for determination of antibody titre of the infected pigs. The capsid protein (Cap) of PCV2d was produced in Saccharomyces cerevisiae cells and the capsid protein was purified by affinity chromatography. This recombinant protein was used as a coating antigen to develop a cost effective, highly sensitive and specific single serum dilution ELISA. The in-house developed ELISA was optimized to be used in a 1:200 single serum dilution. The developed ELISA along with a commercial ELISA kit were compared with a sensitive immuno-peroxidase assay (IPMA) by receiver-operating characteristics (ROC) test. Our results showed that the developed single serum dilution ELISA had a higher sensitivity and specificity in comparison to the commercial ELISA. The area under the ROC curve (AUC) also confirmed that the developed ELISA had a better overall diagnostic performance in comparison to the commercial ELISA kit.
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Acknowledgements
The authors are grateful to the Director, ICAR-Indian Veterinary Research Institute and Director of Research (Veterinary), AAU, Khanapara for providing the facilities to carry out the research work.
Funding
S.D. acknowledges the Department of Biotechnology, Government of India for providing the research support (Grant Order No. BT/PR33050/ADV/90/277/2019). No funding was received to assist with the preparation of this manuscript.
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Conceptualization, S.D. and C.M.M.; methodology, R.Y.V., D.P., A.S., R.M., V.M., S.D., C.M.M., and N.N.B.; formal analysis, S.D., C.M.M. and L.B.; investigation S.D., L.B., R.M. and A.S.; writing—original draft preparation, R.Y.V., L.B., S.D., D.P. and R.M.; writing—review and editing S.D., C.M.M., L.B., N.N.B. and D.P.; all authors have read and agreed to the published version of the manuscript.
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The study involved blood sampling from animals. The study protocol was assessed and approved by the Institute Animal Ethics Committee of Assam Agricultural University, Khanapara, Guwahati vide Approval No. 770/GO/Re/S/03/CPCSEA/FVSc/AAU/IAEC/20–21/876 dated 31.07.2021. Owners of the animals provided their verbal informed consent for animal blood sampling. Collection of blood samples was carried out by trained veterinarians adhering to the regulations and guidelines on animal husbandry and welfare.
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Yadhapura Venkatesh, R., Dey, S., Buragohain, L. et al. Development of recombinant capsid protein-based single serum dilution ELISA for sero-detection of porcine circovirus type 2 infection in pigs. Vet Res Commun 48, 1149–1159 (2024). https://doi.org/10.1007/s11259-024-10299-y
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DOI: https://doi.org/10.1007/s11259-024-10299-y