Development and evaluation of a rapid multianalyte particle-based flow cytometric assay for the quantification of meningococcal serogroup B-specific IgM antibodies in sera for nonculture case confirmation

Abstract

Serogroup-specific antibody has been shown to be present in the sera of patients recovering from meningococcal disease, and thus the detection of such antibodies may aid in the confirmation of disease. There are currently no standard methods for measuring meningococcal serogroup B-specific antibody in sera. Here, we report the development of a microsphere-based immunoassay which utilizes colominic acid from Escherichia coli 07:K1 (L):NM to detect immunoglobulin M directed against serogroup B polysaccharide. The serogroup B assay was incorporated into a multiplex assay which also detects serogroup-specific immunoglobulin M for meningococcal serogroups A, C, Y and W-135. Using the method of crossstandardization, serogroup B-specific immunoglobulin M concentrations were assigned to the standard serum CDC 1992. The assay is able to detect increases in specific immunoglobulin M concentrations from acute to convalescent phase serum from serogroup B cases, and can be utilized in conjunction with the previously developed tetraplex immunoglobulin G detection assay for serogroups A, C, Y and W-135.

Age-Related Immunity to Meningococcal Serogroup C Vaccination: An Increase in the Persistence of IgG2 Correlates with a Decrease in the Avidity of IgG

“…Meningococcal serogroup A and C-specific IgG1, IgG2 and MenC PS-specific IgM antibodies were quantified using a fluorescent-bead-based multiplex immunoassay (MIA) as published before [14], [15]. Standardized reference serum CDC 1992 was used in this assay (NIBSC, Potters Bar, UK).…”

Inadequacy of Colominic Acid as an Absorbent Intended To Facilitate Use of Complement-Preserved Baby Rabbit Serum in theNeisseria meningitidisSerogroup B Serum Bactericidal Antibody Assay

“…Using a subset of samples (n ϭ 36), anti-colominic acid immunoglobulin M (IgM) concentrations were determined using a bead-based assay (17). The methodology was adapted to detect anti-colominic acid IgG concentrations by the inclusion of R-phycoerythrin-conjugated goat anti-human IgG Fc␥ fragment (Jackson ImmunoResearch Laboratories, PA).…”

Multiplexed Immunoassays

“…At present, the most successful and robust multiplexing technology is Luminex xMAP platform, which combines advanced fluidics, optics, and digital signal processing with proprietary microsphere technology to deliver multiplexed assay capabilities. Importantly, Luminex is an open technology and numerous companies have marketed the Luminex system, including Bio-Rad, Qiagen, Invitrogen, and Millipore ect.. And a steadily growing list of ready-to-use multiplexed immunoassays have also been provided by these companies for applications in biomarker discovery , autoimmune disease diagnostics [77][78][79][80] , infectious disease diagnostics [81][82][83][84][85][86][87][88][89][90][91][92][93] , neurological diseases [94][95][96][97][98][99][100][101] , HLA testing [102][103][104] , and drug discovery 105,106 ( Table 4).…”

“…Moreover, encoded microsphere based multiplexed immunoassay technology could improve diagnostics of infectious diseases by enabling the simultaneous detection of antibodies or antigens to multiple infectious pathogens, such as human immunodeficiency virus (HIV), the Hepatitis A, B, C virus, Mycobacterium tuberculosis, as well as a large number of other viral, bacterial and parasitic pathogens. [81][82][83][84][85][86][87][88][89][90][91][92][93] FDA-approved assays for infectious diseases (e.g. EBV, HSV, MMRV, Syphilis, and ToRC assays on BioPlex TM 2200 or Multi-Lyte TM ) are already available on market.…”