Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay

J. Mertsola, R. S. Munford, O. Ramilo, X. Saez-Llorens, M. M. Mustafa, G. H. McCracken, E. J. Hansen

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Three monoclonal antibody (MAb)-based immunoassays were developed for specific detection of Haemophilus influenzae type b (Hib) lipooligosaccharide (LOS). (i) Hib LOS was captured onto microtiter plates by polyclonal Hib-directed antibodies and detected with MAbs in the oligosaccharide component of Hib LOS in an enzyme-linked immunosorbent assay. (ii) The high affinity of polymyxin B for lipid A was used to bind Hib LOS to microtiter wells, and the oligosaccharide-specific MAbs were used as the detection system in the polymyxin B-MAb assay. (iii) Hib LOS solubilized in detergent was captured by MAbs, and the immobilized LOS was detected with a chromogenic Limulus amebocyte lysate method in the immunolimulus assay. Endotoxin concentrations were measured in in vitro samples and cerebrospinal fluid samples from rabbits with experimental Hib meningitis. The results were compared with those obtained with the standard chromogenic Limulus amebocyte lysate assay. There were significant correlations between the results of all four assays. These new immunoassays provide methods for specific detection of Hib LOS in laboratory fluids and in research involving quantification of Hib endotoxin in experimental animal models.

Original languageEnglish (US)
Pages (from-to)2700-2706
Number of pages7
JournalJournal of Clinical Microbiology
Volume28
Issue number12
StatePublished - 1990

Fingerprint

Haemophilus influenzae type b
Immunoassay
Horseshoe Crabs
Polymyxin B
Oligosaccharides
Endotoxins
Monoclonal Antibodies
lipid-linked oligosaccharides
Lipid A
Meningitis
Detergents
Cerebrospinal Fluid
Animal Models
Enzyme-Linked Immunosorbent Assay
Rabbits
Antibodies

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Mertsola, J., Munford, R. S., Ramilo, O., Saez-Llorens, X., Mustafa, M. M., McCracken, G. H., & Hansen, E. J. (1990). Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay. Journal of Clinical Microbiology, 28(12), 2700-2706.

Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay. / Mertsola, J.; Munford, R. S.; Ramilo, O.; Saez-Llorens, X.; Mustafa, M. M.; McCracken, G. H.; Hansen, E. J.

In: Journal of Clinical Microbiology, Vol. 28, No. 12, 1990, p. 2700-2706.

Research output: Contribution to journalArticle

Mertsola, J, Munford, RS, Ramilo, O, Saez-Llorens, X, Mustafa, MM, McCracken, GH & Hansen, EJ 1990, 'Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay', Journal of Clinical Microbiology, vol. 28, no. 12, pp. 2700-2706.
Mertsola J, Munford RS, Ramilo O, Saez-Llorens X, Mustafa MM, McCracken GH et al. Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay. Journal of Clinical Microbiology. 1990;28(12):2700-2706.
Mertsola, J. ; Munford, R. S. ; Ramilo, O. ; Saez-Llorens, X. ; Mustafa, M. M. ; McCracken, G. H. ; Hansen, E. J. / Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay. In: Journal of Clinical Microbiology. 1990 ; Vol. 28, No. 12. pp. 2700-2706.
@article{05aa8f689f3d48e18bd9eabd0be23bae,
title = "Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay",
abstract = "Three monoclonal antibody (MAb)-based immunoassays were developed for specific detection of Haemophilus influenzae type b (Hib) lipooligosaccharide (LOS). (i) Hib LOS was captured onto microtiter plates by polyclonal Hib-directed antibodies and detected with MAbs in the oligosaccharide component of Hib LOS in an enzyme-linked immunosorbent assay. (ii) The high affinity of polymyxin B for lipid A was used to bind Hib LOS to microtiter wells, and the oligosaccharide-specific MAbs were used as the detection system in the polymyxin B-MAb assay. (iii) Hib LOS solubilized in detergent was captured by MAbs, and the immobilized LOS was detected with a chromogenic Limulus amebocyte lysate method in the immunolimulus assay. Endotoxin concentrations were measured in in vitro samples and cerebrospinal fluid samples from rabbits with experimental Hib meningitis. The results were compared with those obtained with the standard chromogenic Limulus amebocyte lysate assay. There were significant correlations between the results of all four assays. These new immunoassays provide methods for specific detection of Hib LOS in laboratory fluids and in research involving quantification of Hib endotoxin in experimental animal models.",
author = "J. Mertsola and Munford, {R. S.} and O. Ramilo and X. Saez-Llorens and Mustafa, {M. M.} and McCracken, {G. H.} and Hansen, {E. J.}",
year = "1990",
language = "English (US)",
volume = "28",
pages = "2700--2706",
journal = "Journal of Clinical Microbiology",
issn = "0095-1137",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - Specific detection of Haemophilus influenzae type b lipooligosaccharide by immunoassay

AU - Mertsola, J.

AU - Munford, R. S.

AU - Ramilo, O.

AU - Saez-Llorens, X.

AU - Mustafa, M. M.

AU - McCracken, G. H.

AU - Hansen, E. J.

PY - 1990

Y1 - 1990

N2 - Three monoclonal antibody (MAb)-based immunoassays were developed for specific detection of Haemophilus influenzae type b (Hib) lipooligosaccharide (LOS). (i) Hib LOS was captured onto microtiter plates by polyclonal Hib-directed antibodies and detected with MAbs in the oligosaccharide component of Hib LOS in an enzyme-linked immunosorbent assay. (ii) The high affinity of polymyxin B for lipid A was used to bind Hib LOS to microtiter wells, and the oligosaccharide-specific MAbs were used as the detection system in the polymyxin B-MAb assay. (iii) Hib LOS solubilized in detergent was captured by MAbs, and the immobilized LOS was detected with a chromogenic Limulus amebocyte lysate method in the immunolimulus assay. Endotoxin concentrations were measured in in vitro samples and cerebrospinal fluid samples from rabbits with experimental Hib meningitis. The results were compared with those obtained with the standard chromogenic Limulus amebocyte lysate assay. There were significant correlations between the results of all four assays. These new immunoassays provide methods for specific detection of Hib LOS in laboratory fluids and in research involving quantification of Hib endotoxin in experimental animal models.

AB - Three monoclonal antibody (MAb)-based immunoassays were developed for specific detection of Haemophilus influenzae type b (Hib) lipooligosaccharide (LOS). (i) Hib LOS was captured onto microtiter plates by polyclonal Hib-directed antibodies and detected with MAbs in the oligosaccharide component of Hib LOS in an enzyme-linked immunosorbent assay. (ii) The high affinity of polymyxin B for lipid A was used to bind Hib LOS to microtiter wells, and the oligosaccharide-specific MAbs were used as the detection system in the polymyxin B-MAb assay. (iii) Hib LOS solubilized in detergent was captured by MAbs, and the immobilized LOS was detected with a chromogenic Limulus amebocyte lysate method in the immunolimulus assay. Endotoxin concentrations were measured in in vitro samples and cerebrospinal fluid samples from rabbits with experimental Hib meningitis. The results were compared with those obtained with the standard chromogenic Limulus amebocyte lysate assay. There were significant correlations between the results of all four assays. These new immunoassays provide methods for specific detection of Hib LOS in laboratory fluids and in research involving quantification of Hib endotoxin in experimental animal models.

UR - http://www.scopus.com/inward/record.url?scp=0025204093&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025204093&partnerID=8YFLogxK

M3 - Article

C2 - 2280001

AN - SCOPUS:0025204093

VL - 28

SP - 2700

EP - 2706

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

SN - 0095-1137

IS - 12

ER -