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Published on: October 12, 2013
Bovine herpesvirus type 1 glycoprotein C expression in MDBK cells and its reactivity in enzyme-linked immunosorbent
P K Gupta1, M Saini, S K Bandyopadhyay
1National Biotechnology Centre, Indian Veterinary Research Institute, India.
Insights
Bovine herpesvirus type 1 glycoprotein C (gC) was successfully expressed in MDBK cells. This method provides a comparable alternative to purified antigen for detecting BHV-1 antibodies via ELISA.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Glycoprotein C (gC) is a key surface protein of bovine herpesvirus type 1 (BHV-1), crucial for viral infection and a primary target for immune responses.
- Understanding gC expression and its role in immunity is vital for developing effective diagnostics and vaccines against BHV-1.
Purpose of the Study:
- To express the bovine herpesvirus type 1 (BHV-1) glycoprotein C (gC) in MDBK cells.
- To validate the use of these gC-expressing cells as an antigen in enzyme-linked immunosorbent assays (ELISA) for detecting BHV-1 antibodies.
Main Methods:
- Transfection of MDBK cells with the pRSV-gC plasmid containing the BHV-1 gC gene.
- Detection of gC expression using SDS-PAGE and immunoblot analysis.
- Confirmation of gC gene transcription via RT-PCR.
- Utilizing gC-expressing MDBK cells as antigen in ELISA for field sera analysis.
Main Results:
- Successful expression of BHV-1 gC in transfected MDBK cells was confirmed.
- RT-PCR verified the transcription of the gC gene in these cells.
- ELISA results using gC-expressing cells as antigen showed high comparability (92.44%) with results using purified BHV-1 antigen.
Conclusions:
- MDBK cells can be effectively utilized for the expression of BHV-1 gC.
- gC-expressing MDBK cells serve as a viable and comparable antigen for BHV-1 antibody detection in ELISA, offering a potential alternative to purified viral antigens.
Abstract:
Glycoprotein C (gC) of bovine herpesvirus type 1 (BHV-1) is a major viral glycoprotein expressed at high level on the surface of infected cells and on the virion envelope. This glycoprotein is also a major target of immune response at both humoral and cellular levels. The plasmid pRSV-gC having complete coding gene for BHV-1 gC was transfected into MDBK cells and the expression of gC in these cells was detected by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) and immunoblot analysis. Transcription of the gC gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) of total RNA isolated from transfected cells. MDBK cells expressing BHV-1 gC were used as antigen in enzyme-linked immunosorbent assay (ELISA) of antibodies against BHV-1 in field sera. The results were found comparable (92.44%) with those obtained with BHV-1 purified antigen.
