Prokaryotic expression of a VP1 polypeptide antigen for diagnosis by a human parvovirus B19 antibody enzyme immunoassay.
To produce parvovirus B19 antigen for diagnostic purposes, partially overlapping segments covering the genes encoding the viral structural proteins VP1 and VP2 were cloned into expression vectors. The constructs were induced in Escherichia coli, resulting in the expression of beta-galactosidase fusion proteins. In immunoblotting experiments with sera from patients with erythema infectiosum, immunoglobulin G (IgG) and IgM antibodies bound to a single polypeptide of 235 amino acids at the N terminus of VP1. The DNA fragment encoding this polypeptide was amplified by the polymerase chain reaction and cloned into an expression vector. The viral capsid antigen expressed in E. coli was purified by preparative agarose gel electrophoresis and used in IgG and IgM solid-phase enzyme immunoassays. Comparison with reference gamma- and mu-capture radioimmunoassays using whole virus antigen showed that these antibody tests are suitable for the serodiagnosis of human infections caused by parvovirus B19.
Söderlund
M
M
Department of Virology, University of Helsinki, Finland.
Brown
K E
KE
Meurman
O
O
Hedman
K
K
D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics; D002213:Capsid / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D003001:Cloning, Molecular; D016731:Erythema Infectiosum / Q000175:diagnosis*; D004926:Escherichia coli / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D007124:Immunoenzyme Techniques; D016732:Parvovirus B19, Human / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins