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Clinical and Diagnostic Laboratory Immunology, July 2002, p. 872-876, Vol. 9, No. 4
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.4.872-876.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Comparison of a Multiplex Flow Cytometric Assay with Enzyme-Linked Immunosorbent Assay for Quantitation of Antibodies to Tetanus, Diphtheria, and Haemophilus influenzae Type b

Jerry W. Pickering,1* Thomas B. Martins,1 M. Carl Schroder,1 and Harry R. Hill1,2

Associated Regional and University Pathologists, Institute for Clinical and Experimental Pathology,1 Department of Pathology, Pediatrics and Medicine, University of Utah School of Medicine, Salt Lake City, Utah 841082

Received 11 February 2002/ Returned for modification 20 March 2002/ Accepted 24 April 2002

We developed a multiplexed indirect immunofluorescence assay for antibodies to Haemophilus influenza type b (Hib) polysaccharide and the toxoids of Clostridium tetani (Tet) and Corynebacterium diphtheriae (Dip) based on the Luminex multiple-analyte profiling system. A pooled serum standard was calibrated against World Health Organization standards for Dip and Tet and an international standard for Hib. The multiplexed Luminex assay was compared to individual enzyme-linked immunosorbent assays (ELISAs) for the same analytes. By both methods, 75 (92.6%) of 81 of random serum samples had protective levels of antibody to Tet (>=0.1 IU/ml). For Dip, 81.5% of the samples had protective antibody levels (>=0.1 IU/ml) by ELISA and 80.2% had protective antibody levels by Luminex. Protective levels (>=1.0 µg/ml) of antibody to Hib were found in 45.0% of the samples tested by ELISA and in 39.0% of the samples tested by Luminex. The correlations (R2) between ELISA and Luminex of the 81 samples were 0.96, 0.96, and 0.91 for Tet, Dip, and Hib, respectively. There was also similar agreement between Luminex and ELISA for sera collected before and 1 month after Tet, Dip, and Hib vaccine administration. Both methods detected strong postvaccination responses. The Luminex method is an attractive alternative to ELISA since it reduces labor and reagent costs, as well as assay time.

* Corresponding author. Mailing address: ARUP Institute, 500 Chipeta Way, Salt Lake City, UT 84108. Phone: (801) 583-2787. Fax: (801) 583-2712. E-mail: pickerjw{at}aruplab.com.

Clinical and Diagnostic Laboratory Immunology, July 2002, p. 872-876, Vol. 9, No. 4
1071-412X/02/$04.00+0     DOI: 10.1128/CDLI.9.4.872-876.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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