Diagnostics of Pogosta disease: antigenic properties and evaluation of Sindbis virus IgM and IgG enzyme immunoassays.
Sindbis virus (SINV) is a mosquito-borne causative agent of a fever-rash arthritis, Pogosta disease, as verified recently by virus isolation from acutely ill patients. Pogosta disease occurs annually, but it emerges as unique epidemics every 7 years in Finland; over 10,000 patient samples have been analyzed for SINV antibodies, with over 2000 diagnosed acute SINV infections. However, the performance of these serological tests with a large number of samples has not been described before. The aim of the present study was to characterize and evaluate methods developed for the serodiagnostics of SINV infection, suitable for large sample numbers, and to examine the protein-specific responses to the antigen used. We developed SINV IgM and IgG enzyme immunoassays (EIA) using highly purified SINV. The EIAs were compared to hemagglutination inhibition (HI) and neutralization tests. We studied paired samples from 46 acutely ill patients taken at approximately 2-week intervals, with a verified SINV infection confirmed by a fourfold rise in HI antibody titer. The assay cut-off values and specificity were determined with confirmed negative sera. Protein-specific antibody responses were examined with immunoblot assay. The optical density values of the EIAs correlated well with the HI titers. The sensitivities of the IgM and IgG EIAs were 97.6% and 100%, and specificities were 95.2% and 97.6%, respectively. We consider that a verified serological diagnosis of acute SINV infection requires (1) in addition to a positive IgM result at least a fourfold increase in HI (or IgG) titer between paired sera or (2) a positive IgM result and a negative/borderline IgG result (which excludes old immunity) and specific reaction in HI. Both E1 and E2 glycoproteins of SINV were shown to be recognized by IgM and IgG antibodies early in infection.
Manni
Tytti
T
Department of Virology, Haartman Institute, Faculty of Medicine, University of Helsinki, Finland.
Kurkela
Satu
S
Vaheri
Antti
A
Vapalahti
Olli
O
D018354:Alphavirus Infections / Q000175:diagnosis* / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000276:immunology; D001170:Arthritis, Infectious / Q000821:virology*; D006386:Hemagglutination Tests; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000276:immunology*; D007075:Immunoglobulin M / Q000276:immunology*; D009500:Neutralization Tests; D012680:Sensitivity and Specificity; D012845:Sindbis Virus / Q000276:immunology*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M