Specific detection of Haemophilus influenzae type b lipooligosaccharide by a polymyxin B monoclonal antibody assay

Jussi Mertsola, Octavio Ramilo, Xavier Sáez-Llorens, Mark S. Hanson, George H. McCracken, Eric J. Hansen

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

A monoclonal antibody (MAb)-based enzyme immunoassay was developed for detection of Haemophilus influenzae type b (Hib) lipooligosaccharides (LOS). The high affinity of polymyxin B for lipid A was used to bind the Hib LOS to microtiter wells. The immobilized LOS was detected with MAbs directed against the oligosaccharide component of Hib endotoxin. Hib LOS concentrations were measured in in vitro samples and in cerebrospinal fluid (CSF) samples obtained from rabbits with experimental Hib meningitis. The sensitivity of the assay was 1 ng LOS/ml sample and the results obtained with this assay correlated significantly with those obtained with the standard Limulus amebocyte lysate assay. This new assay provides a method for specific detection of Hib LOS in CSF samples and in aqueous laboratory fluids. This general methodology should also be useful for experimental research involving specific LPS/LOS molecules.

Original languageEnglish (US)
Pages (from-to)219-226
Number of pages8
JournalJournal of Immunological Methods
Volume122
Issue number2
DOIs
StatePublished - Sep 1 1989

Keywords

  • Endotoxin
  • Haemophilus influenzae type b
  • Limulus amebocyte lysate method
  • Lipooligosaccharide
  • Monoclonal antibody
  • Polymyxin B

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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