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.
Laboratorio di Immunochimica e Sierologia
Sperimentale,
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.
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