An ELISA employing a Haemophilus influenzae type b oligosaccharide-human serum albumin conjugate correlates with the radioantigen binding assay.
An ELISA measuring total Ig antibodies to the capsular polysaccharide of Haemophilus influenzae type b (HbPs) in human sera using an antigen composed of Haemophilus b oligosaccharides conjugated to human serum albumin (HbO-HA) was shown to have an excellent correlation to the radioantigen binding assay (RABA). When 214 sera with different anti-HbPs levels were assayed for total Ig by HbO-HA ELISA and by RABA the correlation coefficient was 0.917 and the paired t test p value was 0.575. Use of competitive ELISA employing soluble HbPs, HbO-HA and human albumin as competitors, showed that the HbO-HA ELISA was specific for antibodies to HbPs. The HbO-HA ELISA yielded reproducible results both within and between laboratories. The HbO-HA ELISA can also be used to determine the isotype of anti-HbPs antibodies by using isotype specific enzyme conjugates. The sum of the IgG, IgA and IgM HbO-HA ELISA results had excellent correlation to the RABA results (correlation coefficient = 0.976). Thus, the HbO-HA ELISA can be substituted for the classical RABA and also be utilized for quantitating the isotype of the anti-HbPs antibodies.
Phipps
D C
DC
Department of Immunobiologic Laboratory Service, Praxis Biologics, Inc., Rochester, NY 14623.
West
J
J
Eby
R
R
Koster
M
M
Madore
D V
DV
Quataert
S A
SA
D000328:Adult; D000907:Antibodies, Bacterial / Q000097:blood* / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D016667:Bacterial Capsules; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology*; D001667:Binding, Competitive / Q000276:immunology; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D018073:Haemophilus Vaccines; D006193:Haemophilus influenzae / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M / Q000032:analysis; D011135:Polysaccharides, Bacterial / Q000008:administration & dosage / Q000276:immunology*; D011869:Radioligand Assay / Q000379:methods; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000276:immunology*; D014611:Vaccination
D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D018073:Haemophilus Vaccines; C044237:Haemophilus influenzae type b polysaccharide vaccine; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D011135:Polysaccharides, Bacterial; D012709:Serum Albumin