Comparison of three ELISA protocols to measure antibody responses elicited against serogroup C meningococcal polysaccharide in mouse, monkey and human sera

In this study we compared the following ELISA protocols to measure antibody levels against serogroup C meningococcal polysaccharide: a traditional protocol using poly- l-Lysine mixed with the polysaccharide as coating antigen, a second protocol coating with a mixture of methylated human serum albumi...

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Bibliographic Details
Published in:Journal of microbiological methods 2006-04, Vol.65 (1), p.135-143
Main Authors: Guirola, María, Carmenate, Tania, Menéndez, Tamara, Álvarez, Anabel, González, Sonia, Guillén, Gerardo
Format: Article
Language:English
Subjects:
Animals
Antibodies, Bacterial - biosynthesis
Antibodies, Bacterial - blood
Bacterial Capsules - immunology
Bacteriology
Biological and medical sciences
Capsular polysaccharide
Cercopithecus aethiops
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Female
Fundamental and applied biological sciences. Psychology
Genetics
Humans
Immune response
Meningitis, Meningococcal - immunology
Meningitis, Meningococcal - microbiology
Meningitis, Meningococcal - pathology
Meningococcal Vaccines - immunology
Mice
Mice, Inbred BALB C
Microbiology
Neisseria meningitidis
Neisseria meningitidis, Serogroup C - immunology
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Polysaccharides, Bacterial - immunology
Statistics, Nonparametric
Vaccines, Conjugate - immunology
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Summary:In this study we compared the following ELISA protocols to measure antibody levels against serogroup C meningococcal polysaccharide: a traditional protocol using poly- l-Lysine mixed with the polysaccharide as coating antigen, a second protocol coating with a mixture of methylated human serum albumin with the C polysaccharide, a modified protocol coating with derivatized polysaccharide and a modification to the last one, specifically without adding ammonium thiocyanate to the sample buffer. Serum bactericidal activity of mouse, monkey and human sera were measured and correlation coefficients were calculated. For all serum types the modified ELISA protocol showed the highest correlation coefficients while the traditional protocol showed the lower ones. We demonstrated that the traditional protocol measures non-specific antibodies to the C polysaccharide, because no differences were detected between pre-immune and post-immune human sera ( P > 0.05), while the modified protocol detected the highest difference ( P < 0.01).
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2005.06.015