Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus
Effective surveillance of viral disease in fish populations is critical for disease control and the sustainable development of global aquaculture. Here, we evaluated the application and performance of pooled serum samples using an indirect ELISA based on recombinant segment 4 protein to assess farm-...
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MDPI AG
2025-06-01
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| Online Access: | https://www.mdpi.com/1999-4915/17/7/877 |
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| author | Jidapa Yamkasem Puntanat Tattiyapong Ian A. Gardner Win Surachetpong |
| author_facet | Jidapa Yamkasem Puntanat Tattiyapong Ian A. Gardner Win Surachetpong |
| author_sort | Jidapa Yamkasem |
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| description | Effective surveillance of viral disease in fish populations is critical for disease control and the sustainable development of global aquaculture. Here, we evaluated the application and performance of pooled serum samples using an indirect ELISA based on recombinant segment 4 protein to assess farm-level immunity in tilapia infected with Tilapia lake virus (TiLV). The TiLV-S4 ELISA was developed using a recombinant nucleoprotein (segment 4) antigen, optimized through checkerboard titration, and validated for repeatability and reproducibility, with intra- and inter-assay coefficients of variation below 10%. A pooling strategy was used to combine multiple serum samples before testing for the presence of TiLV-specific antibodies using an enzyme-linked immunosorbent assay (ELISA). Our results showed that pooling five serum samples was effective for detecting TiLV-specific antibodies, particularly when multiple seropositive individuals were presented in the pool, supporting its application for population-level surveillance. However, ELISA sensitivity may be reduced when only one seropositive sample is included in the pool, due to the dilution effects. Despite this limitation, pooled testing yielded a high proportion of positive results, suggesting similar detection performance in many cases. Overall, the pooling strategy provides a cost-effective and time-efficient approach for large-scale monitoring of immune status in tilapia populations. |
| format | Article |
| id | doaj-art-531def8ea7454f349982b89c6bbbba74 |
| institution | DOAJ |
| issn | 1999-4915 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
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| series | Viruses |
| spelling | doaj-art-531def8ea7454f349982b89c6bbbba742025-08-20T02:47:21ZengMDPI AGViruses1999-49152025-06-0117787710.3390/v17070877Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake VirusJidapa Yamkasem0Puntanat Tattiyapong1Ian A. Gardner2Win Surachetpong3Department of Veterinary Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, ThailandDepartment of Veterinary Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, ThailandAtlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PE C1A4P3, CanadaDepartment of Veterinary Microbiology and Immunology, Faculty of Veterinary Medicine, Kasetsart University, Bangkok 10900, ThailandEffective surveillance of viral disease in fish populations is critical for disease control and the sustainable development of global aquaculture. Here, we evaluated the application and performance of pooled serum samples using an indirect ELISA based on recombinant segment 4 protein to assess farm-level immunity in tilapia infected with Tilapia lake virus (TiLV). The TiLV-S4 ELISA was developed using a recombinant nucleoprotein (segment 4) antigen, optimized through checkerboard titration, and validated for repeatability and reproducibility, with intra- and inter-assay coefficients of variation below 10%. A pooling strategy was used to combine multiple serum samples before testing for the presence of TiLV-specific antibodies using an enzyme-linked immunosorbent assay (ELISA). Our results showed that pooling five serum samples was effective for detecting TiLV-specific antibodies, particularly when multiple seropositive individuals were presented in the pool, supporting its application for population-level surveillance. However, ELISA sensitivity may be reduced when only one seropositive sample is included in the pool, due to the dilution effects. Despite this limitation, pooled testing yielded a high proportion of positive results, suggesting similar detection performance in many cases. Overall, the pooling strategy provides a cost-effective and time-efficient approach for large-scale monitoring of immune status in tilapia populations.https://www.mdpi.com/1999-4915/17/7/877TiLVimmunityELISApoolingserum samplestilapia |
| spellingShingle | Jidapa Yamkasem Puntanat Tattiyapong Ian A. Gardner Win Surachetpong Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus Viruses TiLV immunity ELISA pooling serum samples tilapia |
| title | Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus |
| title_full | Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus |
| title_fullStr | Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus |
| title_full_unstemmed | Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus |
| title_short | Assessment and Performance of Pooled Serum Samples for Monitoring Farm-Level Immunity in Tilapia Infected with Tilapia Lake Virus |
| title_sort | assessment and performance of pooled serum samples for monitoring farm level immunity in tilapia infected with tilapia lake virus |
| topic | TiLV immunity ELISA pooling serum samples tilapia |
| url | https://www.mdpi.com/1999-4915/17/7/877 |
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