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Updated: Jun 26, 2026
An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis
Published on: October 4, 2018
Replication of classical infectious bursal disease virus in the chicken embryo related cell line
1Departamento de Apoio, Producao e Saude Animal, da Faculdade de Odontologia, Curso de Medicina Veterinaria, Rua Clovis Pestana, 793, Universidade Estadual Paulista, Aracatuba, SP, CEP 16.050-680, Brazil.
Insights
Diagnosing Infectious Bursal Disease Virus (IBDV) is challenging. This study shows Chicken Embryo Related (CER) cells can propagate IBDV, with detection possible via DAS-ELISA and RT-PCR as early as 6 hours post-infection.
Area of Science:
- Veterinary Virology
- Cell Biology
Background:
- Infectious Bursal Disease (IBD) is a highly contagious viral disease affecting poultry.
- Current diagnostic methods for IBDV are time-consuming and expensive, involving virus isolation and histopathology.
- There is a need for efficient and rapid diagnostic techniques for IBDV detection.
Purpose of the Study:
- To evaluate the efficacy of Chicken Embryo Related (CER) cell line for the propagation of classical Infectious Bursal Disease Virus (IBDV).
- To compare the diagnostic performance of DAS-ELISA, immunoperoxidase, and RT-PCR for early IBDV detection in infected cell cultures.
Main Methods:
- Classical IBDV (Lukert strain) was passaged three times in CER cell monolayers.
- Infected CER cells were analyzed at 6, 12, 24, and 48 hours post-infection (p.i.).
- Detection methods included cytopathic effect observation, Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA), immunoperoxidase staining, and Reverse Transcription Polymerase Chain Reaction (RT-PCR).
Main Results:
- Cytopathic effects in CER cells were most evident at 12 h.p.i. but visible from 6 h.p.i.
- DAS-ELISA showed maximum discrimination between infected and uninfected cells at 12 h.p.i. (0.597 +/-0.02 vs 0.010 +/-0.01).
- RT-PCR successfully detected viral RNA from 6 h.p.i. using primers targeting the VP2 region.
Conclusions:
- The CER cell line is suitable for the propagation of classical IBDV.
- DAS-ELISA and RT-PCR are effective methods for the rapid detection of IBDV in cell cultures.
- Early detection of IBDV is achievable within 6 hours post-infection using molecular techniques like RT-PCR.
Abstract:
Infectious bursal disease (IBD) is an acute, highly contagious viral disease. The diagnosis of IBD depends on time-consuming and costly procedures, like virus isolation on chick embryos and histopathological examination. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), immunoperoxidase and reverse transcription polymerase chain reaction (RT-PCR) were applied in this study to detect classical IBD virus (IBDV) after three blind passages of the Lukert strain on chicken embryo related (CER) cell monolayer after different periods of infection: 6, 12, 24 and 48h. Cytophatic effects were most evident 12 h post-infection (p.i.) but were observed at 6 h p.i. The maximum discrimination between IBDV-infected and uninfected cell suspensions obtained by the use of DAS-ELISA for virus detection corresponded to 0.597 +/-0.02 and 0.010 +/-0.01 after 12 h p.i., respectively. The RT-PCR was performed using the set of primers A3.1 and A3.2 to amplify the VP2 region of the IBDV genome. This molecular technique demonstrated that from 6h p.i., it was possible to detect the viral RNA. The results show that the CER cell line can be used for classical IBDV propagation, confirmed by the DAS-ELISA, immunoperoxidase and RT-PCR assay.
