Pedv infection generates conformation-specific antibodies that can be effectively detected by a cell-based elisa
Journal
Viruses
Journal Volume
13
Journal Issue
2
Date Issued
2021
Author(s)
Abstract
Porcine epidemic diarrhea virus (PEDV) is a coronavirus that causes serious and highly con-tagious enteric disease in swine worldwide. In this study, we constructed a recombinant baculovirus (S-Bac) expressing full-length spike protein of the virulent epidemic genotype 2b (G2b) PEDV strain for serological studies of infected pigs. We found that most spike-specific antibodies produced upon PEDV infection in pigs are conformation-specific and they could be detected on S-Bac-infected insect cells by immunofluorescent assay, but they were insensitive to Western blot analysis, the typical method for antiserum analysis. These results indicated that spike conformation is crucial for serum recognition. Since it is difficult to purify trimeric spike membrane protein for conventional enzyme-linked immunosorbent assay (ELISA), we used S-Bac to generate a novel cell-based ELISA for convenient PEDV detection. We analyzed 100 pig serum samples, and our cell-based ELISA exhibited a sensitivity of 100%, a specificity of 97%, and almost perfect agreement [Cohen’s kappa coefficient value (κ) = 0.98] with immunocytochemical staining results. Our cell-based ELISA rapidly presented antigen for proper detection of conformation-specific antibodies, making PEDV detection more convenient, and it will be useful for detecting many viral diseases in the future. ? 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Subjects
coronavirus spike glycoprotein; recombinant protein; virus antibody; virus antigen; animal; Baculoviridae; blood; Chlorocebus aethiops; Coronavirus infection; enzyme linked immunosorbent assay; immunology; pig; Porcine epidemic diarrhea virus; Spodoptera; swine disease; Vero cell line; veterinary medicine; virology; Animals; Antibodies, Viral; Antigens, Viral; Baculoviridae; Chlorocebus aethiops; Coronavirus Infections; Enzyme-Linked Immunosorbent Assay; Porcine epidemic diarrhea virus; Recombinant Proteins; Spike Glycoprotein, Coronavirus; Spodoptera; Swine; Swine Diseases; Vero Cells
SDGs
Other Subjects
antiserum; cell surface protein; membrane protein; protein antibody; virus antibody; virus spike protein; vitronectin; coronavirus spike glycoprotein; recombinant protein; virus antibody; virus antigen; animal cell; animal experiment; animal model; animal tissue; Article; cell culture; cell membrane; cell surface; controlled study; diagnostic test accuracy study; enzyme linked immunosorbent assay; ID50 (median infectious dose); immunocytochemistry; immunofluorescence assay; nonhuman; plasmid; polyacrylamide gel electrophoresis; porcine epidemic diarrhea; Porcine epidemic diarrhea virus; protein conformation; protein expression; sensitivity and specificity; Sf21 cell line; Spodoptera frugiperda; Vero cell line; virus load; virus recombinant; virus strain; Western blotting; animal; Baculoviridae; blood; Chlorocebus aethiops; Coronavirus infection; immunology; pig; Spodoptera; swine disease; veterinary medicine; virology; Animals; Antibodies, Viral; Antigens, Viral; Baculoviridae; Chlorocebus aethiops; Coronavirus Infections; Enzyme-Linked Immunosorbent Assay; Porcine epidemic diarrhea virus; Recombinant Proteins; Spike Glycoprotein, Coronavirus; Spodoptera; Swine; Swine Diseases; Vero Cells
Type
journal article