title: Development of a bead immunoassay to measure Vi polysaccharide-specific serum IgG after vaccination with the Salmonella enterica serovar Typhi Vi polysaccharide.
abstract: Vi polysaccharide from Salmonella enterica serotype Typhi is used as one of the available vaccines to prevent typhoid fever. Measurement of Vi-specific serum antibodies after vaccination with Vi polysaccharide by enzyme-linked immunosorbent assay (ELISA) may be complicated due to poor binding of the Vi polysaccharide to ELISA plates resulting in poor reproducibility of measured antibody responses. We chemically conjugated Vi polysaccharide to fluorescent beads and performed studies to determine if a bead-based immunoassay provided a reproducible method to measure vaccine-induced anti-Vi serum IgG antibodies. Compared to ELISA, the Vi bead immunoassay had a lower background and therefore a greater signal-to-noise ratio. The Vi bead immunoassay was used to evaluate serum anti-Vi IgG in 996 subjects from the city of Kolkata, India, before and after vaccination. Due to the location being one where Salmonella serotype Typhi is endemic, approximately 45% of the subjects had protective levels of anti-Vi serum IgG (i.e., 1 microg/ml anti-Vi IgG) before vaccination, and nearly 98% of the subjects had protective levels of anti-Vi serum IgG after vaccination. Our results demonstrate that a bead-based immunoassay provides an effective, reproducible method to measure serum anti-Vi IgG responses before and after vaccination with the Vi polysaccharide vaccine.
authors.affiliation: Department of Pathology, Duke University Medical Center, Durham, NC 27710, USA. herman.staats@duke.edu
authors.forename: Herman F
authors.initials: HF
authors.lastname: Staats
authors.forename: Shaun M
authors.initials: SM
authors.lastname: Kirwan
authors.forename: Carol C
authors.initials: CC
authors.lastname: Whisnant
authors.forename: James L
authors.initials: JL
authors.lastname: Stephenson
authors.forename: Diane K
authors.initials: DK
authors.lastname: Wagener
authors.forename: Partha P
authors.initials: PP
authors.lastname: Majumder
mesh_terms: D000907:Antibodies, Bacterial / Q000032:analysis / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood*; D007194:India; D011135:Polysaccharides, Bacterial / Q000276:immunology*; D022562:Salmonella Vaccines / Q000276:immunology*; D012485:Salmonella typhi / Q000276:immunology; D014435:Typhoid Fever / Q000517:prevention & control; D014436:Typhoid-Paratyphoid Vaccines / Q000276:immunology*
chemical_list: D000907:Antibodies, Bacterial; D007074:Immunoglobulin G; D011135:Polysaccharides, Bacterial; D022562:Salmonella Vaccines; D014436:Typhoid-Paratyphoid Vaccines; C054154:capsular polysaccharide, Salmonella