Development of a peptide-based enzyme-linked immunosorbent assay (ELISA) for serosurveillance of tilapia lake virus in farmed tilapia | |
| Author | Gaurav, Veer |
| Call Number | AIT Thesis no.AQ-23-03 |
| Subject(s) | Fishes--Diseases--Prevention Peptides--Synthesis Tilapia |
| Note | A thesis submitted in partial fulfillment of the requirements for the degree of Master of Engineering in Aquaculture and Aquatic Resources Management |
| Publisher | Asian Institute of Technology |
| Abstract | Disease caused by tilapia lake virus (TiLV) is a significant threat to farmed tilapia globally. Previous studies have reported that TiLV vaccines elicit specific antibodies that protect fish from viral infection. Currently, the measurement of anti-TiLV antibodies by enzyme-linked immunosorbent assay (ELISA) relies on the use of viral particles (VP), which require a cell culture platform that is unavailable in many laboratories across Asia and Africa. In this study, we investigated whether synthetic peptides P1 and P2 derived from predicted B cell epitopes of TiLV segments 5 and 6, respectively, can replace viral particles for ELISA. Optimization of coated antigen and serum dilutions demonstrated that 2μg peptide/well and a dilution of 256-fold gave the best results for ELISA assay. The optimized ELISA protocols do not cross-react with sera obtained from fish immunized with Aeromonas veronii and Streptococcus agalactiae. P1, P2, a mixture of P1+P2, and VP were then assayed in parallel with sera samples from naive, TiLV-exposed tilapia, and TiLV-vaccinated tilapia. The results showed that P1, P2, and P1+P2 had similar performance as VP, suggesting that these synthetic peptides can replace VP for measuring the antibody response post-vaccination and TiLV serosurveillance. This finding enables many aquatic animal laboratories without accessible cell culture systems to use synthetic peptides for serosurveillance and evaluation of fish immune response post-vaccination. |
| Year | 2023 |
| Type | Thesis |
| School | School of Environment, Resources, and Development |
| Department | Department of Food, Agriculture and Natural Resources (Former title: Department of Food Agriculture, and BioResources (DFAB)) |
| Academic Program/FoS | Aquaculture and Aquatic Resources Management (AQ) |
| Chairperson(s) | Ha, Thanh Dong;Salin, Krishna R. (Co-Chairperson) |
| Examination Committee(s) | Loc Thai Nguyen;Lumpan Poolsawat |
| Scholarship Donor(s) | AIT Scholarships and Student Assistantship |
| Degree | Thesis (M. Eng.) - Asian Institute of Technology, 2023 |