GLYCOPROTEINS OF THE EMBRYONATED EGG INFECTED WITH PR8 INFLUENZA A VIRUS.

GLYCOPROTEINS OF THE EMBRYONATED EGG INFECTED WITH PR8 INFLUENZA A VIRUS

Detection of Anti-Arboviral Immunoglobulin G by Using a Monoclonal Antibody-Based Capture Enzyme-Linked Immunosorbent Assay

Monoclonal antibody (MAb)-based capture enzyme-linked immunosorbent assays (ELISAs) for the detection of anti-arboviral immunoglobulin G (IgG ELISAs) were developed for a comprehensive array of medically important arboviruses from the Alphavirus, Flavivirus, and Bunyavirus genera. Tests were optimized and standardized so that maximum homology could be maintained among working parameters for the different viral agents, enabling a wide range of viruses to be easily tested for at one time. MAbs were screened for suitability as capture vehicles for antigens from the three genera. The final test configuration utilized group-reactive MAbs eastern equine encephalitis virus 1A4B-6, dengue 2 virus 4G2, and La Crosse encephalitis virus 10G5.4 to capture the specific inactivated viral antigens. Serum IgG was detected by using alkaline phosphatase-conjugated anti-human IgG (Fc portion). A dilution of 1:400 was chosen as the universal screening serum dilution, with endpoint titrations of serum samples testing positive eliminating occasional false-positive results. IgG ELISA results correlated with those of the standard plaque-reduction neutralization assays. As expected, some test cross-reactivity was encountered within the individual genera, and tests were interpreted within the context of these reactions. The tests were standardized for laboratory diagnosis of arboviral infections, with the intent that they be used in tandem with the corresponding IgM antibody-capture ELISAs

Standardization of Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assays for Routine Diagnosis of Arboviral Infections

Immunoglobulin M antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA) is a rapid and versatile diagnostic method that readily permits the combination of multiple assays. Test consolidation is especially important for arthropod-borne viruses (arboviruses) which belong to at least three virus families: the Togaviridae, Flaviviridae, and Bunyaviridae. Using prototype viruses from each of these families and a panel of well-characterized human sera, we have evaluated and standardized a combined MAC-ELISA capable of identifying virus infections caused by members of each virus family. Furthermore, by grouping antigens geographically and utilizing known serological cross-reactivities, we have reduced the number of antigens necessary for testing, while maintaining adequate detection sensitivity. We have determined that a 1:400 serum dilution is most appropriate for screening antiviral antibody, using a positive-to-negative ratio of ≥2.0 as a positive cutoff value. With a blind-coded human serum panel, this combined MAC-ELISA was shown to have test sensitivity and specificity that correlated well with those of other serological techniques

Key words: flaviviruses/neutralization/antigenic relationships Antigenic Relationships between Flaviviruses as Determined by Cross-neutralization Tests with Polyclonal Antisera

The recently established virus family Flaviviridae contains at least 68 recognized members. Sixty-six of these viruses were tested by cross-neutralization in cell cultures. Flaviviruses were separated into eight complexes [tick-borne encephalitis (12 viruses), Rio Bravo (six), Japanese encephalitis (10), Tyuleniy (three), Ntaya (five), Uganda S (four), dengue (four) and Modoc (five)] containing 49 viruses; 17 other viruses were not sufficiently related to warrant inclusion in any of these complexes

Antigenic relationships among rhabdoviruses from vertebrates and hematophagous arthropods

Centers for Disease Control. Public Health Service. US Department ofHealth and Human Services. Center for Infectious Diseases. Division ofVector-Borne Diseases. Arbovirus Reference Branch. Fort Collins, Colo, USA.Centers for Disease Control. Public Health Service. US Department ofHealth and Human Services. Center for Infectious Diseases. Division ofVector-Borne Diseases. Arbovirus Reference Branch. Fort Collins, Colo, USACenters for Disease Control. Public Health Service. US Department ofHealth and Human Services. Center for Infectious Diseases. Division of Vector-Borne Diseases. Arbovirus Reference Branch. Fort Collins, Colo, USA / Pasteur Institute. Dakar, Senegal.Pasteur Institute. Dakar, Senegal.Yale University School of Medicine. Departmentof Epidemiology and Public Health. Yale Arbovirus Research Unit. New Haven, CT, USA.Yale University School of Medicine. Departmentof Epidemiology and Public Health. Yale Arbovirus Research Unit. New Haven, CT, USA.Ministério da Saúde. Fundação Serviços de Saúde Pública. Instituto Evandro Chagas. Belém, PA, Brasil.Ministério da Saúde. Fundação Serviços de Saúde Pública. Instituto Evandro Chagas. Belém, PA, Brasil.Commonwealth Scientific and Industrial Research Organization. Division of Tropical Animal Science. Indooroopilly, Queensland, Australia