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 optimiz...

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Bibliographic Details
Published in:Journal of clinical microbiology Vol. 38; no. 5; p. 1827
Main Authors: Johnson, A J, Martin, D A, Karabatsos, N, Roehrig, J T
Format: Journal Article
Language:English
Published: United States 01.05.2000
Subjects:
Alphavirus Infections - diagnosis
Antibodies, Monoclonal
Antibodies, Viral - blood
Antigens, Viral - immunology
Arbovirus Infections - blood
Arbovirus Infections - diagnosis
Arbovirus Infections - immunology
Bunyaviridae Infections - diagnosis
Centers for Disease Control and Prevention (U.S.)
Cross Reactions
Dengue - diagnosis
Diagnosis, Differential
Encephalitis, California - diagnosis
Encephalomyelitis, Equine - diagnosis
Enzyme-Linked Immunosorbent Assay - methods
Flavivirus Infections - diagnosis
Humans
Immunoglobulin G - blood
La Crosse virus
Reproducibility of Results
United States
Viral Plaque Assay
United States
ISSN:0095-1137
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Summary: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.
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ISSN:0095-1137
DOI:10.1128/JCM.38.5.1827-1831.2000