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Link to original content: https://pubmed.ncbi.nlm.nih.gov/15879016/
Duplex microsphere-based immunoassay for detection of anti-West Nile virus and anti-St. Louis encephalitis virus immunoglobulin m antibodies - PubMed Skip to main page content
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. 2005 May;12(5):566-74.
doi: 10.1128/CDLI.12.5.566-574.2005.

Duplex microsphere-based immunoassay for detection of anti-West Nile virus and anti-St. Louis encephalitis virus immunoglobulin m antibodies

Alison J Johnson  1 , Amanda J Noga, Olga Kosoy, Robert S Lanciotti, Alicia A Johnson, Brad J Biggerstaff
Affiliations

Duplex microsphere-based immunoassay for detection of anti-West Nile virus and anti-St. Louis encephalitis virus immunoglobulin m antibodies

Alison J Johnson et al. Clin Diagn Lab Immunol. 2005 May.

Abstract

West Nile (WN) virus was introduced into the United States in 1999, when the first human cases of WN fever and encephalitis appeared in New York City. From there, the virus has spread throughout North America, in some areas cocirculating with the related flavivirus St. Louis encephalitis (SLE) virus. Public health laboratories currently use an immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) as a primary test for human serodiagnosis, followed by a confirmatory plaque-reduction neutralization test (PRNT). The MAC-ELISAs take 2 days to perform; therefore there is a need for a more rapid test. This report describes a duplex microsphere-based immunoassay (MIA) that shortens the test processing time to about 4.5 h. The assay employs two sets of microspheres coupled to a single flavivirus group-reactive antibody, which are used to capture the WN and SLE viral antigens independently. Immunoglobulin G-depleted serum is concurrently assayed for IgM antibodies to each of the viral antigens. The results are standardized and classified by using quadratic discriminant analysis so that a single result, anti-WN IgM-positive, anti-SLE IgM-positive, negative, or nonspecific, can be determined. The duplex MIA results compared favorably to those of the plaque-reduction neutralization test and MAC-ELISA. The assay proved to be reproducible, produced accurate classifications as to the infecting virus, and was specific.

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Figures

FIG. 1.
FIG. 1.
Specimens used in the WN/SLE duplex MIA study. Sources and set names and subset names are listed, with numbers of samples shown in parentheses.
FIG. 2.
FIG. 2.
Quadratic discriminant analysis (QDA) classification of test sera. The line represents the classification rule such that the antivirus IgM classifications by the QDA analysis are as annotated. The symbols represent the true classification by PRNT (or MAC-ELISA for some negative samples).
FIG. 3.
FIG. 3.
Comparison of true classifications and MAC-ELISA results for the samples used in the QDA. The equivocal zone is defined as 2 ≤ P/N < 3.
FIG. 4.
FIG. 4.
Classification probability surface. The contour and three-dimensional plots represent the classification probabilities computed on a grid over the range of observed data. The discrimination line in Fig. 2 is the bottom of the trough on the surface. The contour plot is an interpretation of the surface, where lines closer together indicate a steeper grade on the surface.
See this image and copyright information in PMC

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