A new enzyme-linked immunosorbent assay for serological diagnosis of seal parapoxvirus infection in marine mammals
Pubblicato online: 01 mar 2022
Pagine: 43 - 52
Ricevuto: 06 set 2021
Accettato: 24 gen 2022
DOI: https://doi.org/10.2478/jvetres-2022-0005
Parole chiave
© 2022 Y. Badr et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.
Introduction
Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection.
Material and Methods
The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (
Results
Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal.
Conclusion
The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.