Development, Interlaboratory Evaluations, and Application of a Simple, High-Throughput Shigella Serum Bactericidal Assay
PublisherAmerican Society for Microbiology
MetadataShow full item record
AbstractShigella is an important cause of diarrhea worldwide, with serotypes Shigella flexneri 2a, S. flexneri 3a, and Shigella sonnei demonstrating epidemiological prevalence. Many development efforts are focused on Shigella lipopolysaccharide (LPS)-based vaccines, as O antigen-specific conjugate vaccines are immunogenic and efficacious. Immunization with Shigella vaccines containing LPS can elicit antibodies capable of killing Shigella in a serotype-specific manner. Thus, to facilitate Shigella vaccine development, we have developed a serum bactericidal assay (SBA) specific for three Shigella serotypes that measures killing of target bacteria at multiple serum dilutions and in the presence of exogenous complement. The SBA has a high analytical throughput and uses simple technologies and readily available reagents. The SBA was characterized with human sera with bactericidal antibodies against S. flexneri 2a, S. flexneri 3a, and S. sonnei. Purified LPS of a homologous serotype, but not a heterologous serotype, inhibited bacterial killing. Assessment of precision found median intra-assay precision to be 13.3% and median interassay precision to be 19 to 30% for the three serotypes. The SBA is linear, with slight deviations for samples with low (~40) killing indices. The SBA was sensitive enough to allow about 100-fold predilution of serum samples. Repeat assays yielded results with less than 2-fold deviations, indicating the robustness of the assay. Assay results from four different laboratories were highly comparable when normalized with a reference serum. The Shigella SBA, combined with a reference serum, should facilitate the development of Shigella vaccines across the field. Copyright 2018 Nahm et al.
Identifier to cite or link to this itemhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85054015474&doi=10.1128%2fmSphere.00146-18&partnerID=40&md5=6e98df66a07bf771e4443a1238b7fd8b; http://hdl.handle.net/10713/9416
- Monoclonal Antibodies to Shigella Lipopolysaccharide Are Useful for Vaccine Production.
- Authors: Lin J, Smith MA, Benjamin WH Jr, Kaminski RW, Wenzel H, Nahm MH
- Issue date: 2016 Aug
- Functional and Antigen-Specific Serum Antibody Levels as Correlates of Protection against Shigellosis in a Controlled Human Challenge Study.
- Authors: Shimanovich AA, Buskirk AD, Heine SJ, Blackwelder WC, Wahid R, Kotloff KL, Pasetti MF
- Issue date: 2017 Feb
- Development of a high-throughput method to evaluate serum bactericidal activity using bacterial ATP measurement as survival readout.
- Authors: Necchi F, Saul A, Rondini S
- Issue date: 2017
- Shigella-Specific Immune Profiles Induced after Parenteral Immunization or Oral Challenge with Either Shigella flexneri 2a or Shigella sonnei.
- Authors: Clarkson KA, Porter CK, Talaat KR, Frenck RW Jr, Alaimo C, Martin P, Bourgeois AL, Kaminski RW
- Issue date: 2021 Aug 25
- Quantitative analysis of IgG class and subclass and IgA serum response to Shigella sonnei and Shigella flexneri 2a polysaccharides following vaccination with Shigella conjugate vaccines.
- Authors: Robin G, Keisari Y, Slepon R, Ashkenazi S, Cohen D
- Issue date: 1999 Aug 6