development of a multi-plex electrochemiluminescent assay for the detection of serum antibody responses to meningococcal conjugate vaccines发展multi-plex electrochemiluminescent测定血清抗体的检测对脑膜炎球菌结合疫苗的反应.pdfVIP

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development of a multi-plex electrochemiluminescent assay for the detection of serum antibody responses to meningococcal conjugate vaccines发展multi-plex electrochemiluminescent测定血清抗体的检测对脑膜炎球菌结合疫苗的反应.pdf

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development of a multi-plex electrochemiluminescent assay for the detection of serum antibody responses to meningococcal conjugate vaccines发展multi-plex electrochemiluminescent测定血清抗体的检测对脑膜炎球菌结合疫苗的反应

World Journal of Vaccines, 2012, 2, 27-35 27 /10.4236/wjv.2012.21004 Published Online February 2012 (http://www.SciRP.org/journal/wjv) Development of a Multi-Plex Electrochemiluminescent Assay for the Detection of Serum Antibody Responses to Meningococcal Conjugate Vaccines * Lani Indrawati , Jon H. Heinrichs, Emily P. Wen, Julie M. Skinner Vaccines Research, Merck Research Laboratories, West Point, New York, USA. * Email: lani_indrawati@ Received November 10th, 2011; revised December 20th, 2011; accepted January 15th, 2012 ABSTRACT Neisseria meningitidis is a gram negative diplococcal bacterium. Worldwide, N. meningitidis is the leading cause of bacterial meningitis and sepsis, with five serogroups (A, B, C, Y, and W-135) responsible for the majority of the disease. Multivalent (A, C, Y, and W-135) polysaccharide and conjugate vaccines have been licensed in the United States and elsewhere and are widely available. We have developed a multi-plexed electrochemiluminescent assay to quantitate serum antibody responses to meningococcal polysaccharides A, C, Y, and W-135 to allow for rapid evaluation of li- censed and investigational vaccines. A 96-well plate containing a carbon electrode arrayed with polysaccharides A, C, Y, and W-135 on separate spots within each well has been developed for simultaneous detection of polysaccharide- specific antibodies in serum samples from vaccinated individuals. The assay conditions were optimized using the anti- meningococcal serogroup A/C reference serum pool, CDC 1992 (NIBSC 99/706), through evaluation of plate types, coating polysaccharide concentrations, and blocking and serum diluent buffers. Comparison of single and multi-plex assays demonstrated the sensitivity, specificity, and speed of the multi-plex format for the quantification of serum anti- body re