CVI
Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Mariani, M.
Right arrow Articles by Berbers, G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mariani, M.
Right arrow Articles by Berbers, G.

Clinical and Diagnostic Laboratory Immunology, September 1998, p. 667-674, Vol. 5, No. 5
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

A Competitive Enzyme-Linked Immunosorbent Assay for Measuring the Levels of Serum Antibody to Haemophilus influenzae Type b

Massimo Mariani,1,* Enrico Luzzi,1 Daniela Proietti,1 Silvia Mancianti,1 Daniele Casini,1 Paolo Costantino,2 Pieter van Gageldonk,3 and Guy Berbers3

Laboratorio di Immunochimica e Sierologia Sperimentale, Dipartimento Immunologia, Centro Ricerche,1 Laboratorio di Biochimica, Centro Sviluppo Biotecnologie,2 CHIRON S.p.A., Siena, Italy, and Laboratory for Clinical Vaccine Research, RIVM, Bilthoven, The Netherlands3

Received 12 February 1998/Returned for modification 4 May 1998/Accepted 9 June 1998

A competitive ELISA method is described for the measurement of total antibodies to the capsular polysaccharide of Haemophilus influenzae type b (HibCPS) in human sera. The competitive method showed an excellent correlation to the radioantigen binding assay (RABA, or Farr assay) and improved correlation of sera with low titers with respect to the more conventional noncompetitive method. Overestimation of samples in the low concentration range was no longer observed with the competitive ELISA method. The free HibCPS competition allowed us to eliminate the day-to-day background variation typical of some sera; thus, only values representing the true anti-HibCPS response were determined. The use of precoated microplates, which could be stored up to 8 months, greatly improved the speed of the procedure. An overall correlation coefficient of 0.9660 was found when 407 serum samples with a wide variety of anti-HibCPS antibody levels were tested with the competitive ELISA and RABA. The regression line was very close to the ideal line, with a slope of 1.0045 and an intercept of -0.1996. A subset of 96 serum samples representative of all pre- and postimmunization samples was used to compare the competitive ELISA with a previously described ELISA method. The competitive method performed in two laboratories in different countries showed a better correlation with the RABA. The correlation factors were 0.9770 and 0.9816, respectively, while a factor of 0.9547 was found with the previously described noncompetitive procedure, which was better for this method than previously reported (r = 0.917). Therefore, the competitive ELISA is proposed for the assay of anti-HibCPS titers in sera from vaccinated subjects.


* Corresponding author. Mailing address: Laboratorio di Immunochimica e Sierologia Sperimentale, Dipartimento Immunologia, Centro Ricerche CHIRON S.p.A., Via Fiorentina 1, I-53100 Siena, Italy. Phone: 39-577-243227. Fax: 39-577-243317. E-mail: mariani{at}iris02.biocine.it.


Clinical and Diagnostic Laboratory Immunology, September 1998, p. 667-674, Vol. 5, No. 5
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:




Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Antimicrob. Agents Chemother. Clin. Microbiol. Rev. Infect. Immun.
J. Clin. Microbiol. J. Virol. ALL ASM JOURNALS

Copyright © 1998 by the American Society for Microbiology. All rights reserved.