Development of an indirect enzyme-linked immunosorbent assay test for detecting antibodies to chicken astrovirus in chicken sera.
The development of an indirect enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of Group B chicken astrovirus (CAstV) infections is described. The test was based on the use of an affinity-purified capsid antigen, specific to CAstV isolate 11672, produced as a glutathione-S-transferase N-terminal fusion protein by a recombinant baculovirus. Strongly positive ELISA signals were elicited against experimentally produced antisera raised to CAstVs from Group B (subgroups i and ii) but were negative for antisera raised to a Group A CAstV. Using a panel of 240 selected serum samples, 99% agreement was observed when the results obtained by ELISA were compared to those from an indirect immunofluorescence test for CAstV 11672. The ELISA test was applied to 68 serum sets comprising 1864 samples, which were obtained from parent and grandparent flocks originating mainly in the UK. Of the 52 sets containing ELISA-positive samples, 24 sets had >75% samples positive and nine sets had <25% samples positive and were regarded as having high and low seropositivities, respectively. Of the 1864 serum samples tested 1090 (58.5%) were ELISA positive and of these, 234 sera (21.5%) produced strongly positive signals, whereas moderately positive and weakly positive signals were produced by 562 (51.5%) and 294 (27%) sera. When used for flock screening purposes, this ELISA test can be used to (i) investigate the occurrence of first-time CAstV infections of parent flocks during lay and the possible adverse effects caused by vertically transmitted CAstV infections on broiler hatchability and performance and (ii) diagnose Group B CAstV infections within specific pathogen free flocks.
a Veterinary Sciences Division , Agri-Food & Biosciences Institute , Belfast , UK.
A
A
Skibinska
b School of Biological Sciences, Queen's University of Belfast , Belfast , UK.
A
A
Lee
a Veterinary Sciences Division , Agri-Food & Biosciences Institute , Belfast , UK.
M
M
Wylie
a Veterinary Sciences Division , Agri-Food & Biosciences Institute , Belfast , UK.
V J
VJ
Smyth
a Veterinary Sciences Division , Agri-Food & Biosciences Institute , Belfast , UK.
M D
MD
Welsh
a Veterinary Sciences Division , Agri-Food & Biosciences Institute , Belfast , UK.
D
D
Todd
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D019350:Astroviridae Infections / Q000175:diagnosis / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D052676:Avastrovirus / Q000276:immunology* / Q000302:isolation & purification; D016367:Baculoviridae; D036022:Capsid Proteins / Q000276:immunology; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D007106:Immune Sera; D011201:Poultry Diseases / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D011993:Recombinant Fusion Proteins; D013047:Specific Pathogen-Free Organisms
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007106:Immune Sera; D011993:Recombinant Fusion Proteins