Recombinant Expression of Bornavirus P24 Protein for Enzyme-Linked Immunosorbent Assay Development

Document Type : Research Article

Authors

• Seyedeh Narjes Sadat ¹
• Sahar Khalvand ¹
• Behzad Ramezani ²
• Mahdi Habibi-Anbouhi ³
• Fatemeh Kazemi-Lomedasht ¹
• Hajarsadat Ghaderi ¹
• Mahdi Behdani ⁴

¹ Biotechnology Research Center, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran.

² Kawsar Biotechnology Company, Tehran, Iran.

³ National Cell Bank of Iran, Pasteur Institute of Iran, Tehran, Iran.

⁴ Biotechnology Research Center, Venom & Biotherapeutics Molecules Lab, Pasteur Institute of Iran, Tehran, Iran & Zoonoses Research Center, Pasteur Institute of Iran, Amol, Iran.

10.22067/ijvst.2024.84563.1304

Abstract

Borna disease virus (BDV) is a neurotropic, enveloped and ribonucleic acid (RNA) virus. BDV induces persistent neurologic disease in a wide host range included several vertebrate species as well as human. The BDV genome encodes 6 proteins but p24 protein was identified at higher rates than other proteins at BDV-infected tissues. In this study, BDV-p24 protein was constructed and subcloned into expression plasmid pET22. Confirmation of recombinant protein expression was performed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and western blotting. P24 protein was injected into rabbits with the aim of polyclonal antibody production and immunization. Compared to other diagnostic methods, ELISA is a fast method with cost effective and high sensitivity as well as lower probability of contamination. ELISA method was performed to evaluate the infection in laboratory rabbits and retrospective infection was examined in 50 rabbits. The obtained results in this study indicated that the ELISA method based on p24 protein has a high potential to detect BDV infection.

Keywords

• Borna disease virus
• ELISA
• Polyclonal antibody
• Borna-P24 protein
• Diagnostic method

Main Subjects

• Virology

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