Development and evaluation of a tetraplex flow cytometric assay for quantitation of serum antibodies to Neisseria meningitidis serogroups A, C, Y, and W-135.
A rapid and simple method for the simultaneous quantitation of serum immunoglobulin G (IgG) antibodies specific for Neisseria meningitidis serogroups A, C, Y, and W-135 was developed and evaluated. Four bead sets were generated, each conjugated with one of the meningococcal capsular polysaccharides (A, C, Y, or W-135) and serologically assessed by the use of antimeningococcal international reference sera. Cross-reactivity studies demonstrated no inhibition between monoplex and multiplex assays, and the assay was linear over a 24-fold serum dilution range. Inhibition studies demonstrated that the assay is specific, with <25% heterologous inhibition occurring. The assay was also found to have low intra- and interassay variations and limits of detection < or =650 pg/ml. A comparison of the meningococcal bead assay with the standardized meningococcal enzyme-linked immunosorbent assay showed a good correlation between the IgG concentrations obtained by each assay. The tetraplex assay has the potential to be an important addition to the serologic evaluation of meningococcal capsular polysaccharide conjugate vaccines.
Lal
Gouri
G
Health Protection Agency, Vaccine Evaluation Department, Manchester Royal Infirmary, United Kingdom. gouri.lal@hpa.org.uk
Balmer
Paul
P
Joseph
Helen
H
Dawson
Maureen
M
Borrow
Ray
R
D000907:Antibodies, Bacterial / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry / Q000379:methods* / Q000592:standards; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis; D008589:Meningococcal Infections / Q000175:diagnosis* / Q000276:immunology; D008863:Microspheres; D009345:Neisseria meningitidis / Q000276:immunology* / Q000302:isolation & purification; D038381:Neisseria meningitidis, Serogroup A / Q000276:immunology; D038542:Neisseria meningitidis, Serogroup C / Q000276:immunology; D038582:Neisseria meningitidis, Serogroup W-135 / Q000276:immunology; D038581:Neisseria meningitidis, Serogroup Y / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D007074:Immunoglobulin G