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Two enzyme linked immunosorbent assays for detecting antibodies against meningococcal capsular polysaccharides A and C


, : Two enzyme linked immunosorbent assays for detecting antibodies against meningococcal capsular polysaccharides A and C. Journal of Clinical Pathology 47(5): 405-410

Aims - To evaluate two of the recent methods of coating microtiter plates in the enzyme linked immunosorbent assay (ELISA) for detecting human antibodies against meningococcal capsular polysaccharides A and C with a view to validating a specific meningococcal antibody assay for routine clinical use. Methods - Two four-layer ELISA protocols were standardized: one method utilized meningococcal polysaccharides conjugated to poly-L-lysine polypeptide for coating the microtiter plates; another used polysaccharides mixed with methylated human serum albumin (mHSA). Titration curves were plotted for the ELISAs and the squared Pearson correlation coefficient (R-2) was used to determine the degree of accuracy of fit of the curves. Specificity tests were performed by inhibition and adsorption studies. Results - Both methods gave good titration curves with a high R-2 of gt 0.98, indicating a high degree of accuracy in forming the curves. The titration end point after vaccination, obtained by the mHSA method, was 20 times higher, however, than that obtained by the poly-L-lysine method. Specificity tests showed that in the ELISA using poly-saccharide/poly-L-lysine, antibody activity of a pre-vaccination serum sample was inhibited by 37%, and of post-vaccination serum by 50% with 1000-fold excess antigen. Antibody activity (post-vaccination) was reduced by 51% and 59% respectively, adsorption with antigen-coated Sepharose beads or adsorption with suspensions of killed meningococci. In contrast, antibody activity of a pre-vaccination serum was inhibited by 60% and a post-vaccination serum by 90% in ELISA employing polysaccharides mixed with mHSA. Reproducibility was better with the use of methylated human serum albumin than with poly-L-lysine; the former showed intrabatch and interbatch coefficients of variation of 4% and 2%, respectively, compared with 43% (intrabatch) and 16% (interbatch) obtained with the poly-L-lysine. Conclusion - It is concluded that the antibody assay using meningococcal polysaccharides groups A and C mixed with mHSA is much better than that using polysaccharides coupled with poly-L-lysine.

US$19.90

PMID: 8027392

DOI: 10.1136/jcp.47.5.405


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