Expression of PCV2d Capsid Protein in Escherichia coli and its Evaluation for
Serodiagnosis of Porcine Circovirus Infection
Kartik Samanta2, Seema Rani Pegu2, Gyanendra Singh Sengar2, Kartik Samanta1, Aditya Pratap Acharya1 and Hemanta Kumar Maity1
1Department of Avian Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, India
2Department of Animal Health, ICAR-National Research Centre on Pig, Guwahati, Assam, India
*Corresponding Author: Hemanta Kumar Maity, Department of Avian Sciences, West Bengal University of Animal and Fishery Sciences, Kolkata, India.
December 16, 2022; Published: January 12, 2023
The porcine circovirus 2 (PCV2) is one of the most economically significant swine pathogens, causing PCVAD as well as impaired reproductive efficiency, stillbirth, and weight loss in infected pig populations. The PCV2 ORF2 gene encodes capsid protein (cap protein), which has 233 amino acids (AA) in PCV2a/2b and an additional lysine residue at the C-terminus in PCV2d. In this study, we created a PCV2d capsid sequence (ORF2) for expression in Escherichia coli. His tag chromatography method was used to purify the protein, and western blot was used to detect protein expression. In this study, an indirect enzyme-linked immunosorbent assay (I-ELISA) based on the PCV2d ORF2 protein was developed. The best antigen, serum, and goat anti-porcine IgG conjugate dilutions were 1:1500 (3.245 μg/ml), 1:320, and 1:1600, respectively. The estimated cut-off value was 0.9885, and its analytical sensitivity was 1:2400. Except for PCV2a, PCV2b, and PCV2d antisera, the results of specific evaluation revealed that these antisera did not cross-react with any other common porcine viral pathogens, indicating that this method can be used to detect porcine circovirus infection in pig.
Keywords: PCV2d; ORF2; Capsid protein; Expression; E coli
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