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. 1994 May;47(5):405–410. doi: 10.1136/jcp.47.5.405

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

O A Akinwolere 1, D S Kumararatne 1, R Bartlett 1, D M Goodall 1, D Catty 1
PMCID: PMC502014  PMID: 8027392

Abstract

AIMS--To evaluate two of the recent methods of coating microtitre 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 standardised: one method utilised meningococcal polysaccharides conjugated to poly-L-lysine polypeptide for coating the microtitre plates; another used polysaccharides mixed with methylated human serum albumin (mHSA). Titration curves were plotted for the ELISAs and the squared Pearson correlation coefficient (R2) 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 R2 of > 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 polysaccharide/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, by 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.

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Selected References

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  1. Achtman M., Crowe B. A., Olyhoek A., Strittmatter W., Morelli G. Recent results on epidemic meningococcal meningitis. J Med Microbiol. 1988 Jul;26(3):172–177. [PubMed] [Google Scholar]
  2. Anthony B. F., Concepcion N. F., McGeary S. A., Ward J. I., Heiner D. C., Shapshak P., Insel R. A. Immunospecificity and quantitation of an enzyme-linked immunosorbent assay for group B streptococcal antibody. J Clin Microbiol. 1982 Aug;16(2):350–354. doi: 10.1128/jcm.16.2.350-354.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Artenstein M. S., Brandt B. L., Tramont E. C., Branche W. C., Jr, Fleet H. D., Cohen R. L. Serologic studies of meningococcal infection and polysaccharide vaccination. J Infect Dis. 1971 Sep;124(3):277–288. doi: 10.1093/infdis/124.3.277. [DOI] [PubMed] [Google Scholar]
  4. Artenstein M. S., Gold R., Zimmerly J. G., Wyle F. A., Schneider H., Harkins C. Prevention of meningococcal disease by group C polysaccharide vaccine. N Engl J Med. 1970 Feb 19;282(8):417–420. doi: 10.1056/NEJM197002192820803. [DOI] [PubMed] [Google Scholar]
  5. Beuvery E. C., Kayhty M. H., Leussink A. B., Kanhai V. Comparison of radioimmunoassay and enzyme-linked immunosorbent assay in measurement of antibodies to Neisseria meningitidis group A capsular polysaccharide. J Clin Microbiol. 1984 Oct;20(4):672–676. doi: 10.1128/jcm.20.4.672-676.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beuvery E. C., van Delft R. W., Tiesjema R. H., Nagel J. The enzyme-linked immunosorbent assay of meningococcal and some related Escherichia coli polysaccharides. J Biol Stand. 1983 Jul;11(3):195–204. doi: 10.1016/s0092-1157(83)80006-7. [DOI] [PubMed] [Google Scholar]
  7. Brandt B. L., Wyle F. A., Artenstein M. S. A radioactive antigen-binding assay for Neisseria meningitidis polysaccharide antibody. J Immunol. 1972 Apr;108(4):913–920. [PubMed] [Google Scholar]
  8. Butler J. E., Feldbush T. L., McGivern P. L., Stewart N. The enzyme-linked immunosorbent assay (ELISA): a measure of antibody concentration or affinity. Immunochemistry. 1978 Feb;15(2):131–136. doi: 10.1016/0161-5890(78)90053-6. [DOI] [PubMed] [Google Scholar]
  9. Carlone G. M., Frasch C. E., Siber G. R., Quataert S., Gheesling L. L., Turner S. H., Plikaytis B. D., Helsel L. O., DeWitt W. E., Bibb W. F. Multicenter comparison of levels of antibody to the Neisseria meningitidis group A capsular polysaccharide measured by using an enzyme-linked immunosorbent assay. J Clin Microbiol. 1992 Jan;30(1):154–159. doi: 10.1128/jcm.30.1.154-159.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gold R., Lepow M. L., Goldschneider I., Draper T. L., Gotschlich E. C. Clinical evaluation of group A and group C meningococcal polysaccharide vaccines in infants. J Clin Invest. 1975 Dec;56(6):1536–1547. doi: 10.1172/JCI108235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goldschneider I., Gotschlich E. C., Artenstein M. S. Human immunity to the meningococcus. I. The role of humoral antibodies. J Exp Med. 1969 Jun 1;129(6):1307–1326. doi: 10.1084/jem.129.6.1307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gotschlich E. C., Goldschneider I., Artenstein M. S. Human immunity to the meningococcus. IV. Immunogenicity of group A and group C meningococcal polysaccharides in human volunteers. J Exp Med. 1969 Jun 1;129(6):1367–1384. doi: 10.1084/jem.129.6.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hazlewood M., Nusrat R., Kumararatne D. S., Goodall M., Raykundalia C., Wang D. G., Joyce H. J., Milford-Ward A., Forte M., Pahor A. The acquisition of anti-pneumococcal capsular polysaccharide Haemophilus influenzae type b and tetanus toxoid antibodies, with age, in the UK. Clin Exp Immunol. 1993 Aug;93(2):157–164. doi: 10.1111/j.1365-2249.1993.tb07959.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jones D. M. Epidemiology of meningococcal infection in England and Wales. J Med Microbiol. 1988 Jul;26(3):165–168. [PubMed] [Google Scholar]
  15. Jones D. M. Meningococcal vaccines. J Med Microbiol. 1993 Feb;38(2):77–78. doi: 10.1099/00222615-38-2-77. [DOI] [PubMed] [Google Scholar]
  16. Messina J. P., Hickox P. G., Lepow M. L., Pollara B., Venezia R. A. Modification of a direct enzyme-linked immunosorbent assay for the detection of immunoglobulin G and M antibodies to pneumococcal capsular polysaccharide. J Clin Microbiol. 1985 Mar;21(3):390–394. doi: 10.1128/jcm.21.3.390-394.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mohammed I., Onyemelukwe G. C., Obineche E. N., Gupta N., Oyeyinka G. O. Control of epidemic meningococcal meningitis by mass vaccination. II. Persistence of antibody four years after vaccination. J Infect. 1984 Sep;9(2):197–202. doi: 10.1016/s0163-4453(84)91468-3. [DOI] [PubMed] [Google Scholar]
  18. Mäkelä P. H., Käyhty H., Weckström P., Sivonen A., Renkonen O. V. Effect of group-A meningococcal vaccine in army recruits in Finland. Lancet. 1975 Nov 8;2(7941):883–886. doi: 10.1016/s0140-6736(75)92125-x. [DOI] [PubMed] [Google Scholar]
  19. Roberts R. B. The relationship between group A and group C meningococcal polysaccharides and serum opsonins in man. J Exp Med. 1970 Mar 1;131(3):499–513. doi: 10.1084/jem.131.3.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tramont E. C., Artenstein M. S. Latex agglutination test for measurement of antibodies to meningococcal polysaccharides. Infect Immun. 1972 Mar;5(3):346–351. doi: 10.1128/iai.5.3.346-351.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zigterman G. J., Verheul A. F., Ernste E. B., Rombouts R. F., De Reuver M. J., Jansze M., Snippe H., Willers J. M. Measurement of the humoral immune response against Streptococcus pneumoniae type 3 capsular polysaccharide and oligosaccharide containing antigens by ELISA and ELISPOT techniques. J Immunol Methods. 1988 Jan 21;106(1):101–107. doi: 10.1016/0022-1759(88)90277-3. [DOI] [PubMed] [Google Scholar]

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