Enzyme Linked Immunosorbent Assay (ELISA) for the detection of serum antibodies to the inner core lipopolysaccharide of Neisseria meningitidis Group B

We have developed a solid-phase ELISA to study the human immune response to inner core lipopolysaccharide (LPS) of Neisseria meningitidis ( Nm) using structurally defined glycolipids from a genetically defined mutant ( galE) of a serogroup B Nm strain. Previous studies had demonstrated that a galE (...

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Bibliographic Details
Published in:Journal of immunological methods 2000-04, Vol.237 (1), p.73-84
Main Authors: Plested, Joyce S, Gidney, Margaret Anne J, Coull, Philip A, Griffiths, Helen G, Herbert, Mark A, Bird, A.Graham, Richards, James C, Moxon, E.Richard
Format: Article
Language:English
Subjects:
Acute Disease
Adolescent
Adult
Aged
Animals
Antibodies, Bacterial - blood
Antibodies, Monoclonal
Carbohydrate Sequence
Case-Control Studies
Child
Child, Preschool
ELISA
Enzyme-Linked Immunosorbent Assay - methods
Enzyme-Linked Immunosorbent Assay - statistics & numerical data
GalE mutant
Genes, Bacterial
Humans
IgG
IgM
Immunoglobulin G - blood
Immunoglobulin M - blood
Infant
Inner core LPS
lipopolysaccharides
Lipopolysaccharides - chemistry
Lipopolysaccharides - immunology
Meningococcal Infections - immunology
Mice
Middle Aged
Molecular Sequence Data
Mutation
Neisseria meningitidis
Neisseria meningitidis - classification
Neisseria meningitidis - genetics
Neisseria meningitidis - immunology
Reproducibility of Results
Sensitivity and Specificity
Serotyping
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Summary:We have developed a solid-phase ELISA to study the human immune response to inner core lipopolysaccharide (LPS) of Neisseria meningitidis ( Nm) using structurally defined glycolipids from a genetically defined mutant ( galE) of a serogroup B Nm strain. Previous studies had demonstrated that a galE (inner core) LPS epitope is conserved in ∼70% Nm strains and was accessible to antibody in fully encapsulated wild-type Nm strains. A murine monoclonal antibody, MAb B5, raised to a galE mutant of serogroup B Nm strain, immunotype L3 (B.15.P1.7,16) was used to determine the specificity of the inner core LPS ELISA by inhibition studies using purified galE LPS and human sera. The intra-assay coefficient of variation (CV) was 5–6% and inter-assay CV was 19–22%. Using this ELISA, significant differences in the geometric mean titres (GMTs) of naturally occuring serum antibodies (specific to inner core LPS) between healthy adults (18–65 years, N=54) and healthy infants (3–4 months, N=144) of both IgG and IgM classes were found ( P
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(00)00142-3