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Method for simultaneous measurement of antibodies to 23 pneumococcal capsular polysaccharides.

Authors
Biagini-RE; Schlottmann-SA; Sammons-DL; Smith-JP; Snawder-JC; Striley-CAF; MacKenzie-BA; Weissman-DN
Source
Clin Diagn Lab Immunol 2003 Sep; 10(5):744-750
NIOSHTIC No.
20023573
Abstract
We describe a fluorescent covalent microsphere immunoassay (FCMIA) method for the simultaneous (multiplexed) measurement of immunoglobulin G (IgG) antibodies to 23 pneumococcal capsular polysaccharide (PnPS) serotypes present in the pneumococcal polysaccharide vaccine (PPV23) licensed by the Food and Drug Administration, i.e., PnPSs 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F. In addition, the assay incorporates an internal control that allows for contemporaneous evaluation of the effectiveness of pneumococcal cell wall polysaccharide (C-PS) preadsorption and a second control of PnPS 25 (which is not present in any polysaccharide or conjugate vaccine), which can be used to evaluate interassay reproducibility (useful for pre- versus postvaccination studies). The FCMIA was standardized with U.S. reference antipneumococcal serotype standard serum 89S-2. Preadsorption of 89S-2 with each PnPS and C-PS yielded homologous inhibition for serotypes 1, 6B, 9N, 9V, 11A, 12F,14, 15B, 18C, 19A, 19F, 20, 22F, 25, and 33F; heterologous inhibition for serotypes 9V, 10A, 11A, 12F, 15B, 17F, 20, and 23F; and neither homologous nor heterologous inhibition for serotypes 2, 3, 4, and 5. The minimum detectable concentrations for the 24 multiplexed (PnPS and C-PS) FCMIAs ranged from 20 pg/ml for PnPS 3 to 600 pg/ml for PnPS 14. The PnPS FCMIA method has numerous benefits over enzyme-linked immunosorbent assays commonly used to measure anti-PnPS-specific IgG levels, including increased speed, smaller sample volumes, equivalent or better sensitivity, and increased dynamic range.
Keywords
Polysaccharides; Sampling; Vaccines; Immunotoxins; Bioassays; Serology; Proteins; Antibody-response; Immunological-tests; Immunoglobulins; Enzyme-activity; Enzymes; Laboratory-testing; Biological-monitoring
Contact
Raymond E. Biagini, Division of Applied Research and Technology, National Institute for Occupational Safety and Health, MS-C26, 4676 Columbia Parkway, Cincinnati, OH 45226
CODEN
CDIMEN
Publication Date
20030901
Document Type
Journal Article
Email Address
reb4@cdc.gov
Fiscal Year
2003
NTIS Accession No.
NTIS Price
Issue of Publication
5
ISSN
1071-412X
NIOSH Division
HELD; DART
Source Name
Clinical and Diagnostic Laboratory Immunology
State
OH; WV
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