Development of a multiplexed fluorescent immunoassay for the quantitation of antibody responses to four Neisseria meningitidis serogroups.
Neisseria meningitidis is a gram-negative bacterium causing disease world wide with a fatality rate of 5-10%. Five serogroups, A, B, C, Y and W-135 are responsible for virtually all cases of the disease in humans. We have developed a multiplexed assay for the simultaneous quantitation of IgG antibody responses to the four most immunogenic (A, C, Y, and W-135) N. meningitidis serogroups. A simple and less manipulative method was employed for conjugation of the capsular polysaccharide antigens to the microspheres. The multiplex assay compared well with traditional individual ELISAs, but demonstrated greater than 1 log increase in dynamic range and sensitivity. Specificity studies of the multiplex assay showed greater than 95% homologous inhibition and less than 5% heterologous inhibition for all four serogroups. Intra and inter-assay CVs were generally less than 10% and the limit of detection was <600 pg/ml. The multiplexed assay proved to be reproducible as well as specific and sensitive when compared to the standardized ELISAs. Advantages included a greater dynamic range and simultaneous detection of antibody responses to the four serogroups contained in the tetravalent meningococcal polysaccharide vaccine.
Martins
Thomas B
TB
Associated Regional and University Pathologists (ARUP) Institute for Clinical and Experimental Pathology, Salt Lake City, UT 84108, USA. martintb@aruplab.com
Jaskowski
Troy D
TD
Tebo
Anne
A
Hill
Harry R
HR
D000328:Adult; D000907:Antibodies, Bacterial / Q000032:analysis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology*; D008297:Male; D008589:Meningococcal Infections / Q000276:immunology*; D022401:Meningococcal Vaccines / Q000276:immunology; D008863:Microspheres; D009345:Neisseria meningitidis / Q000276:immunology; D011135:Polysaccharides, Bacterial / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D022401:Meningococcal Vaccines; D011135:Polysaccharides, Bacterial