A new enzyme-linked immunosorbent assay for serological diagnosis of seal parapoxvirus infection in marine mammals

• Published in: Journal of veterinary research Vol. 66; no. 1; pp. 43 - 52
• Main Authors: Badr, Yassien, Rahman, Md. Matiur, Ohno, Yoshito, Ishijima, Keita, Maeda, Ken, Kohyama, Kaoru, Kamatari, Yuji O., Shimizu, Kaori, Okada, Ayaka, Inoshima, Yasuo
• Format: Journal Article
• Language: English
• Published: Poland Sciendo 01.03.2022; De Gruyter Poland
• Subjects: Absorbance; aquarium; Aquariums; blood serum; Cloning; Coatings; Delphinapterus leucas; Diagnosis; DNA; Dolphins & porpoises; ELISA; Enzyme-linked immunosorbent assay; Enzymes; Fusion protein; genes; genetic vectors; green fluorescent protein; Infections; Japan; Lagenorhynchus; Lagenorhynchus obliquidens; Lysates; Mammals; Marine mammals; Medical diagnosis; Parapoxvirus; Phoca largha; pinnipeds; Proteins; seals; serodiagnosis; Serology; seroprevalence; Tursiops truncatus; veterinary medicine; Viral envelope proteins; Western blotting; Japan; parapoxvirus; Western blotting; aquarium; pinnipeds; ELISA
• ISSN: 2450-8608; 2450-7393; 2450-8608
• DOI: 10.2478/jvetres-2022-0005

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. 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 ( ), six beluga whales ( ), three Pacific white-sided dolphins ( ), and ten bottlenose dolphins ( ) from an aquarium in Japan were examined using the ELISA. Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal. The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.