E nteric fever, caused by Salmonella enterica serovars Typhi and Paratyphi, causes substantial illness and death globally (1). However, estimating the population-level burden of infection is challenging. Blood culture, the standard for both diagnosis and surveillance, requires microbiological laboratory facilities that are not available in many low-and-middle-income countries. Challenges in accessing blood culture, along with an estimated diagnostics sensitivity of only 60% (2), contribute to chronic underdetection (3).Juba, the capital of South Sudan, experiences a high burden of enteric infections such as cholera and hepatitis E virus (4,5). Enteric fever is a frequently diagnosed etiology of acute fever, but few laboratories have blood culture capacity for confirmation. Consequentially, the population-level burden of enteric fever is unknown.Hemolysin E (HlyE), a pore-forming toxin, is a sensitive and specific serologic marker for diagnosing typhoidal Salmonella (6-10) and is not associated with typhoid carriage (11). New serologic and analytic tools enable measurement of population-level enteric fever incidence from cross-sectional serosurveys using HlyE IgG and IgA (12). We applied those tools to generate population-level enteric fever seroincidence estimates in Juba.
The StudyWe used dried blood spots (DBS) collected for a SARS-CoV-2 serosurvey in Juba, South Sudan, enrolled during August 7-September 20, 2020; enrollment and sampling methods are described elsewhere (13). In brief, 2-stage cluster sampling was used to randomly select households from predefined enumeration units from 6 administrative divisions within and surrounding Juba; all persons >1 year of age and residing for >1 week within the sampled household were eligible to participate. Capillary blood was collected onto Whatman 903 Protein Saver cards (Sigma-Aldrich, https:// www.sigmaaldrich.com), air dried, and transported at ambient temperature to Massachusetts General Hospital (Boston, MA, USA), where they were stored at 4°C. We tested all banked samples collected from participants <25 years of age and a random sample of participants >25 years of age. Younger participants were prioritized because they matched the age distribution of typhoid case data used for the seroincidence estimation ( 12). The study protocol was approved by ethical review boards with the South Sudan Ministry of Health and Massachusetts General Hospital.We used kinetic ELISAs to quantify HlyE IgA and IgG levels in eluted DBS as described (7,11). To estimate seroincidence, we used the antibody dynamics from a longitudinal cohort of 1,420 blood culture-confirmed enteric fever cases (12). In brief, we created a likelihood function for observed crosssectional population antibody response data based on antibody dynamics after blood-culture confirmed infection. We generated joint incidence estimates by