1. Dried Blood Spots for Measuring Vibrio cholerae-specific Immune Responses
- Author
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Anita S, Iyer, Andrew S, Azman, Malika, Bouhenia, Lul O, Deng, Cole P, Anderson, Michael, Graves, Pavol, Kováč, Peng, Xu, Edward T, Ryan, Jason B, Harris, David A, Sack, Francisco J, Luquero, and Daniel T, Leung
- Subjects
Bacterial Diseases ,Infectious Disease Control ,Physiology ,Immunology ,Antibody Response ,Pathology and Laboratory Medicine ,Research and Analysis Methods ,Proof of Concept Study ,Biochemistry ,Microbiology ,Antibodies ,Specimen Handling ,Sudan ,Cholera ,Immune Physiology ,Vibrio Cholerae ,Medicine and Health Sciences ,Humans ,Serologic Tests ,Desiccation ,Enzyme-Linked Immunoassays ,Immunoassays ,Microbial Pathogens ,Immune Response ,Vibrio ,Vaccines ,Immune System Proteins ,Bacteria ,Organisms ,Biology and Life Sciences ,Proteins ,Cholera Vaccines ,Tropical Diseases ,Antibodies, Bacterial ,Body Fluids ,Bacterial Pathogens ,Blood ,Infectious Diseases ,Medical Microbiology ,Immunologic Techniques ,Anatomy ,Pathogens ,Research Article ,Neglected Tropical Diseases - Abstract
Background Vibrio cholerae causes over 2 million cases of cholera and 90,000 deaths each year. Serosurveillance can be a useful tool for estimating the intensity of cholera transmission and prioritizing populations for cholera control interventions. Current methods involving venous blood draws and downstream specimen storage and transport methods pose logistical challenges in most settings where cholera strikes. To overcome these challenges, we developed methods for determining cholera-specific immune responses from dried blood spots (DBS). Methodology/principal findings As conventional vibriocidal assay methods were unsuitable for DBS eluates from filter paper, we adopted a drop-plate culture method. We show that DBS collected from volunteers in South Sudan, and stored for prolonged periods in field conditions, retained functional vibriocidal antibodies, the titers of which correlated with paired serum titers determined by conventional spectrophotometric methods (r = 0.94, p = 0.00012). We also showed that eluates from DBS Serum Separator cards could be used with conventional spectrophotometric vibriocidal methods, and that they correlated with paired serum at a wide range of titers (r = 0.96, p, Author summary Cholera remains a major public health issue among underprivileged populations in the developing world. Current methods of disease surveillance are inadequate for identifying key populations at highest risk of cholera. Serosurveillance can provide accurate measurements of an individual or population’s exposure to cholera infection or oral cholera vaccine (OCV) induced immunity, though they require venous blood draw and stringent processing needs. Dried blood spots (DBS) overcome these challenges, acting as a portable surveillance tool suitable for field use. We developed a drop-plate culture method for evaluating vibriocidal and cholera-specific isotype responses using DBS from OCV-immunized volunteers from South Sudan. Blood equivalent to only two drops were spotted on Whatman Protein Saver (WPS) DBS cards. Vibriocidal titers from WPS eluates determined by drop-plate culture methods correlated well with serum based assays. In addition, by using DBS cards capable of automatic separation of serum from blood, we demonstrate that vibriocidal titers and V. cholerae polysaccharide antibody responses could be measured by conventional spectrophotometric methods and that these responses are stable over a range of storage temperatures. In summary, we show that cholera-specific immune responses can be measured using DBS, providing a potential tool for large-scale serosurveillance field studies for cholera.
- Published
- 2017