CVI Accepts, published online ahead of print on 4 November 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00289-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Evaluation of pneumococcal polysaccharide immunoassays using 22F adsorption step with serum samples from infants vaccinated with conjugate vaccines

Jan T Poolman^*, Carl E Frasch, Helena Käyhty, Pascal Lestrate, Shabir A Madhi, and Isabelle Henckaerts

GlaxoSmithKline Biologicals, Rixensart, Belgium; Frasch Biologics Consulting, Martinsburg, West Virginia, USA; National Institute for Health and Welfare, Helsinki, Finland; Medical Research Council Respiratory and Meningeal Pathogens Research Unit & Department of Science and Technology/National Research Foundation: Vaccine Preventable Diseases, University of the Witswaterand, Johannesburg, South Africa

^* To whom correspondence should be addressed. Email: jan.poolman{at}gskbio.com.

The history of pneumococcal polysaccharide ELISA is characterized by a continuous search for increased specificity. A third generation ELISA using 22F polysaccharide inhibition has increased assay specificity particularly at low antibody concentrations. In the present work, various 22F/non-22F ELISA comparisons were made, involving three different laboratories, including a WHO reference laboratory, and using sera from subjects from different geographic areas immunized with different pneumococcal conjugate vaccines, including the licensed 7-valent Prevenar^TM and 10-valent Synflorix^TM. All comparisons led to the same conclusion that the threshold defined as 0.35 μg/ml for the WHO non-22F ELISA is lower when using any 22F ELISA. Use of highly purified polysaccharides for coating improved further the specificity of the assay. To conclude, we confirm that 22F ELISA can be recommended as a reference method for the determination of antibodies against pneumococcal polysaccharides.