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Clinical and Diagnostic Laboratory Immunology, Jan 1996, 84-88, Vol 3, No. 1
Copyright © 1996 by the American Society for Microbiology. All rights reserved.

Interlaboratory study evaluating quantitation of antibodies to Haemophilus influenzae type b polysaccharide by enzyme-linked immunosorbent assay

DV Madore, P Anderson, BD Baxter, GM Carlone, KM Edwards, RG Hamilton, P Holder, H Kayhty, DC Phipps, CC Peeters, R Schneerson, GR Siber, JI Ward and CE Frasch
Lederle-Praxis Biologicals, West Henrietta, Rochester, New York, USA.

An interlaboratory study was conducted to determine whether an enzyme- linked immunosorbent assay (ELISA) with an antigen preparation composed of various-sized fragments of Haemophilus influenzae type b polysaccharide conjugated to human serum albumin could be standardized across laboratories and whether the ELISA-derived results from different laboratories are equivalent to those obtained by the standard radioactive antigen binding assay (RABA) for quantitation of anti-H, influenzae type b polysaccharide antibodies. Twenty coded human serum samples were quantitated by ELISA in 11 laboratories and by RABA in 5 laboratories. The mean RABA-derived values served as the basis for all comparisons. While the overall correspondence of antibody values between the two methods was good, significant differences were found among some of the 11 ELISA data sets and among the mean RABA values. Seven laboratories generated higher ELISA antibody values for low- titered sera. Four laboratories generated antibody concentrations that were not statistically different between the two assay methods. The results therefore indicate that the ELISA can tolerate substantial variations in protocol, such as the use of different plates and different antibody reagents, without affecting the quantitation of serum antibodies. However, attention should be focused on low-titered sera, as some assay conditions may yield spurious results. This ELISA is a serologic assay which can serve as an alternative to the RABA for quantitation of antibodies to H. influenzae type h polysaccharide.

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