Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysa...

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Bibliographic Details
Published in:mSphere 2018-08, Vol.3 (4)
Main Authors: Pavliakova, Danka, Giardina, Peter C, Moghazeh, Soraya, Sebastian, Shite, Koster, Maya, Pavliak, Viliam, McKeen, Andrew, French, Roger, Jansen, Kathrin U, Pride, Michael
Format: Article
Language:English
Subjects:
Accuracy
Antibodies, Bacterial - blood
Antigens
Capsular polysaccharides
Clinical Science and Epidemiology
Clinical trials
Cross Reactions
Cross-reactivity
Design of experiments
Enzyme-linked immunosorbent assay
Humans
Immunoassay - methods
Immunoglobulin G
Immunoglobulin G - blood
Immunoglobulins
Lysine
Microspheres
Pneumococcal Vaccines - administration & dosage
Pneumococcal Vaccines - immunology
Poly-L-lysine
Polysaccharides, Bacterial - immunology
Quantitation
Reproducibility of Results
Sensitivity and Specificity
Serotypes
Serum - immunology
Standard deviation
Streptococcus infections
Streptococcus pneumoniae
Streptococcus pneumoniae - immunology
Vaccines
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Summary:A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysaccharide (PnPS) serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. The assay uses poly-l-lysine (PLL)-conjugated PnPS, chemically coupled to spectrally distinct Luminex microspheres. Assay validation experiments were performed using residual human serum samples obtained from 13-valent pneumococcal conjugate vaccine (13vPnC) clinical studies. Assay results are expressed as IgG antibody concentrations in micrograms per milliliter using the international reference serum, 007sp. The lower limit of quantitation (LLOQ) for all serotypes covered in the 13-plex dLIA fell within the range of 0.002 to 0.038 µg/ml serum IgG. The difference between the lower limit and upper limit of the assay range was >500-fold for all serotypes, and assay variability was
ISSN:2379-5042
2379-5042
DOI:10.1128/mSphere.00128-18