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MICROBIAL IMMUNOLOGY

A Latex Bead-Based Flow Cytometric Immunoassay Capable of Simultaneous Typing of Multiple Pneumococcal Serotypes (Multibead Assay)

M. K. Park, D. E. Briles, M. H. Nahm
M. K. Park
Departments of Pediatrics and of
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D. E. Briles
Department of Microbiology and Immunology, University of Alabama, Birmingham, Alabama 35294
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M. H. Nahm
Departments of Pediatrics and of
Pathology and Laboratory Medicine, University of Rochester, Rochester, New York 14642, and
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DOI: 10.1128/CDLI.7.3.486-489.2000
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ABSTRACT

A simple and rapid method of simultaneously determining 15Streptococcus pneumoniae serotypes was developed. Fifteen latex beads of different sizes and different red fluorescence levels were coated with 1 of 15 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9N, 9V, 14, 18C, 19A, 19F, 22F, and 23F) of pneumococcal capsular polysaccharide (PS). The bead mixture was incubated with individual pneumococcal lysate, a pool of rabbit antisera capable of binding the 15 serotypes, and fluorescein (green fluorescence)-conjugated anti-rabbit antibody. Bead size, red fluorescence, and green fluorescence were measured in a single flow cytometer run. The green fluorescence of the beads was inhibited only when there was a serotypic match between PS on the bead and PS in the pneumococcal lysate. This method distinguished cross-reactive serotypes and correctly identified the serotypes in 100% of 86 pneumococcal isolates tested.

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A Latex Bead-Based Flow Cytometric Immunoassay Capable of Simultaneous Typing of Multiple Pneumococcal Serotypes (Multibead Assay)
M. K. Park, D. E. Briles, M. H. Nahm
Clinical and Diagnostic Laboratory Immunology May 2000, 7 (3) 486-489; DOI: 10.1128/CDLI.7.3.486-489.2000

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A Latex Bead-Based Flow Cytometric Immunoassay Capable of Simultaneous Typing of Multiple Pneumococcal Serotypes (Multibead Assay)
M. K. Park, D. E. Briles, M. H. Nahm
Clinical and Diagnostic Laboratory Immunology May 2000, 7 (3) 486-489; DOI: 10.1128/CDLI.7.3.486-489.2000
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