Setup of luminescence-based serum bactericidal assay against Salmonella Paratyphi A

Abstract

Increasing awareness of Salmonella Paratyphi A's contribution to enteric fever episodes throughout Asia has led to the development of new S. Paratyphi A vaccines. Assays are needed to measure functional antibodies elicited by the new vaccine candidates to assess their immunogenicity and potential protective capacities. Serum bactericidal assay (SBA) is the method of choice to measure functional antibody titers against various bacterial pathogens, but it is rarely been used for large dataset and clinical samples because it is time consuming and labor-intensive. Recently we developed a high-throughput luminescence-based SBA method, against different pathogens, including Salmonella Typhimurium and Enteritidis, Shigella flexneri serovars 2a and 3a, Shigella sonnei and Neisseria meningitidis. Here we further demonstrated the applicability of such method with invasive isolates of S. Paratyphi A to assess the complement-mediated antibody-dependent killing of both preclinical and clinical standard sera. As already found for other organisms, titers obtained by the luminescence-based SBA strongly correlated with those obtained by the conventional agar plate-based assay. The SBA assay described here is a useful tool for measuring functional antibodies elicited by Salmonella vaccines, with the potential of being applied to immunogenicity assessment in clinical trials.

Keywords: Luminescence; Salmonella Paratyphi A; Serum bactericidal assay.