Children are vulnerable to Taenia solium, a parasitic tapeworm causing cysticercosis (a common cause of epilepsy) and taeniasis. We sought to better understand T. solium prevalence and associated risk factors in Nigeria using data from a 2018 nationally representative household survey estimating HIV incidence. We used a multiplex bead assay to determine the seroprevalence of T. solium IgG antibodies to T24H (cysticercosis) and rES33 (taeniasis) for 32,494 children aged <15 years. Positive antibodies against each antigen were detected in children from all states, with an overall antibody seroprevalence of 6.0% for T24H (range, 1.4-19.3%) and 2.8% for rES33 (range, 0.5-5.3%). Despite state-level variations, overall prevalence for each disease was higher in the north than in the south. We found greater odds of cysticercosis seropositivity among individuals with lower socioeconomic status (aOR = 2.07; 95% CI = 1.53-2.80) when comparing lowest to highest wealth quintile; with increasing age, showing higher odds in children aged 5-9 years (aOR = 2.06; 95% CI = 1.77-2.34) and 10-14 years (aOR = 2.58; 95% CI = 2.22-2.99) compared with children aged <5 years; among households with pig ownership (aOR = 1.35; 95% CI = 1.00-1.83) compared with those without pigs; and among individuals living in rural areas (aOR = 1.26; 95% CI = 1.06-1.51) compared with those in urban areas. For taeniasis, we found significant associations for children in the lowest wealth quintile (aOR = 1.89; 95% CI = 1.30-2.76) and those from pig-owning households (aOR = 1.48; 95% CI = 1.01-2.17). Significant taeniasis prevalence rates were identified even in predominantly Muslim populations. Targeted public health strategies to increase awareness of T. solium infections, highlighting hand hygiene and proper sanitation practices, and improved pig management can help reduce transmission.

Background:Testing blood samples with multiplex bead assays can assess elimination and identify residual foci of transmission for multiple pathogens simultaneously. In Ecuador, onchocerciasis and yaws are presumed eliminated, and the status of trachoma is unknown. We assessed their elimination status by measuring IgG antibodies in children 6-24 months. Methods:In a birth cohort of 404 children measured between 2021-2024 in Esmeraldas province, Ecuador, we tested dried blood spots at ages 6, 9, 12, 18, and 24 months using a multiplex IgG assay that included antigens to Onchocerca volvulus (Ov16), Treponema pallidum (rp17, tmpa), and Chlamydia trachomatis (Pgp3, Ct694). We estimated seroprevalence and incident seroconversion rates across an urban-rural gradient. Results:Of 404 children enrolled, 370 contributed 1,606 samples. Seroprevalence was near zero for onchocerciasis (0.4%, 95% CI: 0.2% to 0.8%) and yaws (0.2%, 95% CI: 0.0% to 0.5%). Conversely, C. trachomatis Pgp3 seroprevalence increased along the urban-rural gradient from Esmeraldas city (3.4%, 95% CI: 1.8% to 5.8%) to remote, river-accessible rural villages (22.4%, 95% CI: 16.0% to 30.1%), and incident seroconversion was common in rural villages (16.1 per 100 child-years, 95% CI: 11.3 to 21.9). Conclusions:Serologic surveillance found no evidence of O. volvulus or T. pallidum transmission, consistent with elimination. High Pgp3 seroconversion rates suggest ongoing C. trachomatis transmission in rural villages. Results highlight the value of integrated serologic surveillance and motivate in-depth trachoma surveillance in Esmeraldas.

The increasing use of serosurveillance for a variety of pathogens calls for systematic studies assessing the stability of total IgG across specimen matrices commonly used in field and laboratory studies. Here, we report results from a two-year study evaluating total IgG stability in six sample matrices (finger-prick dried blood spots, venous blood, and dried serum spots [all on TropBio filter paper], dried blood spots on Whatman filter paper, serum, and plasma). Specimens were stored at -80°C, -20°C, 4°C, 20°C, 30°C, and 37°C to reflect conditions commonly encountered during serosurveys, including short-term exposure to ambient or elevated temperatures during transport, customs delays, or long-term storage before testing. Primary specimens from four donors were tested longitudinally using a multiplex bead assay against a panel of 19 antigens, with parallel evaluations of stored diluted specimens and a batched-testing stability study design. Longitudinal analyses identified storage temperature as the primary driver of signal loss over time, while matrix effects were comparatively small. Storage at -80°C and -20°C were equally robust, with minimal signal loss over two years. Low-positive samples were the most susceptible to elevated temperatures and most likely to lose reactivity. Ambient and elevated temperatures produced measurable changes over two months, but short-term signal loss was limited, supporting specimen transport scenarios. Batched testing provided a suitable alternative design for stability studies, and diluted specimens remained stable for at least one year at 4°C. These findings inform sample storage recommendations, future stability studies, and reliability of multiplex surveillance under real-world conditions.

We evaluated the probability of need for public health action against trachoma in Esmeraldas province, Ecuador. Compared to global references, seroconversion rates to Chlamydia trachomatis Pgp3 in children suggest high probability of action needed in rural villages (91%) and lower in more urban areas (32%), motivating further trachoma assessment.

Objective:To establish baseline seroprevalence of soil-borne, waterborne, and foodborne diseases and to monitor diseases that are eliminated or on the path to elimination in the Paraguayan Chaco. Methods:A total of 1 100 school-age children (6-15 years) were tested in urban and rural schools selected for a cross-cutting population-based survey using a two-stage probabilistic sample design in the three departments of the Paraguayan Chaco. Blood samples were taken on filter paper to measure IgG antibodies using a multiplex bead assay. Data collection was carried out through interviews with parents and caregivers. Access to basic sanitation and improved water was assessed. Differences in pathogen seropositivity and seroprotection were estimated by urban and rural areas. Results:Seroprotection against measles was 62.9% and against rubella was 78.2%. Minimal diphtheria and tetanus seroprotection (≥0.01 IU/ml) was 92.9% and 98.3%, respectively. Seroprotective levels against these four vaccine-preventable diseases significantly decreased with increasing age (p < 0.05). The following pathogens and respective antigens showed significantly higher seroprevalence (p < 0.05) in rural areas compared with urban areas: Cryptosporidium parvum Cp17: 80.4% vs 64.6%, and Cp23: 60.6% vs 44.8%; Giardia lamblia VSP3: 26.9% vs 16.6%; Strongyloides stercoralis NIE: 11.5% vs 4.1%; and Taenia solium T24H: 7.1% vs 1.6%. Seroprevalence for these pathogens was also higher in Indigenous population when compared to non-Indigenous. Basic sanitation conditions showed significant differences (p < 0.05) between rural and urban areas: adobe and soil dwelling floor (65.3% vs 30.2%), use of pit latrine (90.3% vs 44.2%), availability of drainage or septic tank (8.7% vs 55.2%), access to safe water (19.7% vs 44.9%), and water treatment (6.8% vs 32.3%). Conclusions:We identified high exposure to soil-borne, waterborne, and foodborne diseases in rural areas and Indigenous population in the Paraguayan Chaco. Low seroprotection against measles and rubella alerts about the risk of immunity gaps to maintain elimination targets.

Multiplex serological assays simultaneously measure antibodies to multiple antigens, furnishing insights into exposure and susceptibility to several pathogens and cross-pathogen vulnerabilities. Our serosurvey tests dried blood spots from 1292 individuals for IgG antibodies to 35 antigens from 18 pathogens using a multiplex bead assay for vaccine preventable diseases, malaria, SARS-CoV-2, neglected tropical diseases, and enteric pathogens in Mozambique. We produce pathogen-specific seroprevalence estimates and age-seroprevalence curves and identify spatial differences in seroprevalence. Rural clusters have higher odds of seropositivity to most NTDs neglected tropical diseases, Plasmodium falciparum malaria, and enteric pathogens, but lower odds of seropositivity to SARS-CoV-2 and vaccine preventable diseases compared to urban clusters. This co-occurrence identifies clusters with high vulnerability to multiple pathogens. We identify a candidate group of antigens that are correlated with high overall vulnerability. Our results demonstrate a role for multiplex serology in integrated disease surveillance to guide control strategies for individual and co-endemic pathogens.

Objectives Circulating filarial antigen (Ag) is used by elimination programs to monitor lymphatic filariasis (LF) transmission; however, antifilarial antibodies (Ab) may be more sensitive than Ag for detecting LF. Our objectives were to describe Ab seroprevalence, identify risk factors for Ab seropositivity, investigate age-specific associations between Ag and Ab, and evaluate geographic clustering of seropositivity. Methods Community-based serosurveys of participants aged ≥5 years were conducted in 35 primary sampling units (PSUs). Ag-positivity was detected using Alere™ Filariasis Test Strips and Ab-seropositivity using multiplex bead assays. Seroprevalence was adjusted for study design. Results Of 3795 participants (range:5-90 years), adjusted prevalence for Ag, Bm14 Ab, Wb123 Ab, and Bm33 Ab were 3.7% (n=117), 20.3% (n=583), 32.2% (n=987), and 51.0% (n=1659), respectively. Male sex, older age, and residents of suspected hotspots had higher odds of seropositivity to all seromarkers. Seroprevalence was lower in 5-9-year-olds vs ≥10-year-olds (p<0.001). Clustering was significantly higher in households (intra-cluster correlation for Ag:0.45; Bm14 Ab:0.32; Bm33 Ab:0.31; Wb123 Ab:0.29) compared to PSUs or region. Conclusions Abs enabled identification of risk factors for seropositivity and geographical clustering to inform targeted interventions for LF programmes. Further research is needed to define Ab thresholds for active versus past infection and elimination targets.

The multiplex bead assay (MBA) based on Luminex xMAP technology can be used as a tool to measure seroprevalence as part of population immunity evaluations to multiple antigens in large-scale serosurveys. However, multiplexing several antigens presents challenges for quality control (QC) assessments of the data because multiple parameters must be evaluated for each antigen. MBA QC parameters include monitoring bead counts and median fluorescence intensity (MFI) for each antigen in plate wells, and performance of assay controls included on each plate. Analyzing these large datasets to identify plates failing QC standards presents challenges for many laboratories. We developed a novel R Shiny application, shinyMBA, to expedite the MBA QC processes and reduce the risk of user error. The app allows users to rapidly merge multi-plate assay outputs to evaluate bead count, MFI, and performance of assay controls using statistical process control charts for all antigen targets simultaneously. The utility of the shinyMBA application and its various outputs are demonstrated using data from 32 synthetic xPONENT files with 3 multiplex antigens and two population serosurveillance studies that evaluated 1200 and 3871 samples, respectively, for 20 multiplexed antigens. The shinyMBA open-source code is available for download and modification at https://github.com/CDCgov/shinyMBA . Incorporation of shinyMBA into Luminex serosurveillance workflows can vastly improve the speed and accuracy of QC processes.

Yaws is a chronic, relapsing disease of skin, bone, and cartilage caused by Treponema pallidum subsp. pertenue. Yaws was last reported in Nigeria in 1996, although neighboring countries have recently reported cases. We investigated serological evidence for yaws among children aged 0-14 years in Nigeria by measuring antibodies to the treponemal antigens rp17 and TmpA in blood specimens from a 2018 nationally representative HIV survey using a multiplex bead assay. The presence of antibodies to both antigens ("double positive") likely reflects current or recent treponemal infection. Overall, 1.9% (610/31,549) of children had anti-TmpA antibodies, 1.5% (476/31,549) had anti-rp17 antibodies, and 0.1% (39/31,549) were double positive. Among households, 0.5% (84/18,021) had a double-positive child, with a clustering of double-positive children. Although numbers are low, identification of antibodies to both TmpA and rp17 may warrant investigation, including more granular epidemiologic and clinical data, to assess the potential for continuing yaws transmission in Nigerian children.

Serological surveys provide an objective biological measure of population immunity, and tetanus serological surveys can also assess vaccination coverage. We undertook a national assessment of immunity to tetanus and diphtheria among Nigerian children aged <15 years using stored specimens collected during the 2018 Nigeria HIV/AIDS Indicator and Impact Survey, a national cross-sectional household-based survey. We used a validated multiplex bead assay to test for tetanus and diphtheria toxoid-antibodies. In total, 31,456 specimens were tested. Overall, 70.9% and 84.3% of children aged <15 years had at least minimal seroprotection (≥0.01 IU/mL) against tetanus and diphtheria, respectively. Seroprotection was lowest in the north west and north east zones. Factors associated with increased tetanus seroprotection included living in the southern geopolitical zones, urban residence, and higher wealth quintiles (p < 0.001). Full seroprotection (≥0.1 IU/mL) was the same for tetanus (42.2%) and diphtheria (41.7%), while long-term seroprotection (≥1 IU/mL) was 15.1% for tetanus and 6.0% for diphtheria. Full- and long-term seroprotection were higher in boys compared to girls (p < 0.001). Achieving high infant vaccination coverage by targeting specific geographic areas and socio-economic groups and introducing tetanus and diphtheria booster doses in childhood and adolescence are needed to achieve lifelong protection against tetanus and diphtheria and prevent maternal and neonatal tetanus.

Objective:Systematize the experience and identify challenges and lessons learned in the implementation of an initiative for integrated serosurveillance of communicable diseases using a multiplex bead assay in countries of the Americas.Methods:Documents produced in the initiative were compiled and reviewed. These included concept notes, internal working papers, regional meetings reports, and survey protocols from the three participating countries (Mexico, Paraguay, and Brazil) and two additional countries (Guyana and Guatemala) where serology for several communicable diseases was included in neglected tropical diseases surveys. Information was extracted and summarized to describe the experience and the most relevant challenges and lessons learned.Results:Implementing integrated serosurveys requires interprogrammatic and interdisciplinary work teams for the design of survey protocols to respond to key programmatic questions aligned to the needs of the countries. Valid laboratory results are critical and rely on the standardized installment and roll-out of laboratory techniques. Field teams require adequate training and supervision to properly implement survey procedures. The analysis and interpretation of serosurveys results should be antigen-specific, contextualizing the responses for each disease, and triangulated with programmatic and epidemiological data for making decisions tailored to specific population socioeconomic and ecologic contexts.Conclusions:Integrated serosurveillance as a complementary tool for functional epidemiological surveillance systems is feasible to use and key components should be considered: political engagement, technical engagement, and integrated planning. Aspects such as designing the protocol, selecting target populations and diseases, laboratory capacities, anticipating the capacities to analyze and interpret complex data, and how to use it are key.

Diagnostic assays that can simultaneously determine the presence of infection with multiple pathogens are key for diagnosis and surveillance. Current multiplex diagnostic assays are complex and often have limited availability.

BACKGROUND:The drastic decline of Ukraine's immunization coverage since 2009 led to concerns about potential resurgence diphtheria and tetanus, along with other vaccine-preventable diseases. METHODS:To assess population immunity against diphtheria and tetanus, we tested specimens from the serosurvey conducted in 2017 among children born in 2006-2015, the birth cohorts targeted by the nationwide outbreak response immunization following a circulating vaccine-derived poliovirus type 1 outbreak in Zakarpattya province in 2015. We surveyed four regions of Ukraine, using cluster sampling in Zakarpattya, Sumy, and Odessa provinces and simple random sampling in Kyiv City. We tested serum specimens for IgG antibodies against diphtheria and tetanus, using microbead assays (MBA). We estimated seroprevalence and calculated 95% confidence intervals. We also obtained information on the immunization status of surveyed children. RESULTS:Seroprevalence of ≥0.1 IU/mL diphtheria antibodies was <80% in all survey sites (50.0%-79.2%). Seroprevalence of ≥0.1 IU/mL tetanus antibodies was ≥80% in Sumy, Kyiv City, and Odessa (80.2%-89.1%) and 61.6% in Zakarpattya. Across the sites, the proportion of children vaccinated age-appropriately with diphtheria-tetanus-containing vaccines (DTCV) was 28.5%-57.4% among children born in 2006-2010 and 34.1%-54.3% among children born in 2011-2015. The proportion of recipients of <3 DTCV doses increased from 7.1%-16.7% among children born in 2006-2010 to 19.8%-38.6% among children born in 2011-2015, as did the proportion of recipients of zero DTCV doses (2.6%-8.8% versus 8.0%-14.0%, respectively). CONCLUSIONS:Protection against diphtheria among children born in 2006-2015 was suboptimal (<80%), particularly in Zakarpattya. Protection against tetanus was adequate (≥80%) except in Zakarpattya. Diphtheria-tetanus immunization status was suboptimal across all sites. Catch-up vaccination of unvaccinated/under-vaccinated children and other efforts to increase immunization coverage would close these immunity gaps and prevent the resurgence of diphtheria and tetanus in Ukraine, particularly in Zakarpattya.

Trypanosoma cruzi, the causative agent of human Chagas disease, is endemic to the southern region of the United States where it routinely infects many host species. The indoor/outdoor housing configuration used in many non-human primate research and breeding facilities in the southern of the USA provides the opportunity for infection by T. cruzi and thus provides source material for in-depth investigation of host and parasite dynamics in a natural host species under highly controlled and restricted conditions. For cynomolgus macaques housed at such a facility, we used a combination of serial blood quantitative PCR (qPCR) and hemoculture to confirm infection in >92% of seropositive animals, although each method alone failed to detect infection in >20% of cases. Parasite isolates obtained from 43 of the 64 seropositive macaques were of 2 broad genetic types (discrete typing units, (DTU's) I and IV); both within and between these DTU groupings, isolates displayed a wide variation in growth characteristics and virulence, elicited host immune responses, and susceptibility to drug treatment in a mouse model. Likewise, the macaques displayed a diversity in T cell and antibody response profiles that rarely correlated with parasite DTU type, minimum length of infection, or age of the primate. This study reveals the complexity of infection dynamics, parasite phenotypes, and immune response patterns that can occur in a primate group, despite being housed in a uniform environment at a single location, and the limited time period over which the T. cruzi infections were established.

Successful achievement of global targets for elimination of trachoma as a public health problem and eradication of yaws will require control efforts to reach marginalized populations, including refugees. Testing for serologic evidence of transmission of trachoma and yaws in residents of registered camps and a Makeshift Settlement in Cox's Bazar District, Bangladesh, was added to a serosurvey for vaccine-preventable diseases (VPDs) conducted April-May 2018. The survey was primarily designed to estimate remaining immunity gaps for VPDs, including diphtheria, measles, rubella, and polio. Blood specimens from 1- to 14-year-olds from selected households were collected and tested for antibody responses against antigens from Treponema pallidum and Chlamydia trachomatis using a multiplex bead assay to evaluate for serologic evidence of the neglected tropical diseases (NTDs) yaws and trachoma, respectively. The prevalence of antibodies against two C. trachomatis antigens in children ranged from 1.4% to 1.5% for Pgp3 and 2.8% to 7.0% for CT694. The prevalence of antibody responses against both of two treponemal antigens (recombinant protein17 and treponemal membrane protein A) tested was 0% to 0.15% in two camps. The data are suggestive of very low or no transmission of trachoma and yaws, currently or previously, in children resident in these communities. This study illustrates how integrated serologic testing can provide needed data to help NTD programs prioritize limited resources.