Performance of Urine Reagent Strips in Detecting the Presence and Estimating the Prevalence and Intensity of Schistosoma haematobium Infection.

The performance of the urine reagent strips (URS) in detecting the presence and estimating the intensity of Schistosoma haematobium infection was evaluated using urine filtration microscopy as a reference standard. Urine samples collected from 1288 school-age children living in five villages of the Afar and one village in the Gambella Regional States of Ethiopia between October 2021 and April 2022 were examined using urine filtration and URS. The prevalence of S. haematobium infection was 31.6% based on urine filtration and 32.1% using URS. Using results of the urine filtration as a reference, the sensitivity, specificity, negative predictive values, and accuracy of the URS in detecting S. haematobium egg-positive urine specimens were 73.7%, 87.8%, 87.1%, and 82.8%, respectively. Sensitivity increased significantly with an increase in the urine egg count. Specificity was greater in low prevalence settings and among children aged 5-9 years. The level of hematuria detected was trace (19.1%), weak (30.2%), moderate (36.0%), or high (14.7%). The log odds of showing higher-level hematuria significantly increased as the number of egg counts in urine increased. In conclusion, URS remains good in rapidly screening individuals for S. haematobium infection, but the sensitivity of the test could be lower, particularly when the intensity of the infection is light.

Prevalence and factors associated with transmission of schistosomiasis in school-aged children in South Sudan: a cross-sectional study.

Introduction: South Sudan is affected by a high burden of Neglected Tropical Diseases (NTDs). The country is very vulnerable to NTDs due to its favourable tropical climate and multiple risk factors. However, the distribution of the diseases and the populations at risk for the various NTDs is unknown. This paper described the distribution of schistosomiasis in 58 counties and 261 schools in South Sudan. Methods: a descriptive quantitative cross-sectional study of schistosomiasis in 58 counties in 8 states of South Sudan recruited school-aged children. Using different laboratory techniques, the children were tested for Schistosoma mansoni (S. mansoni) and Schistosoma haematobium (S. haematobium). A quantitative descriptive statistical was performed to determine the prevalence rates and the endemicity of schistosomiasis among 13,286 school-aged children. Results: the overall prevalence of S. mansoni and S. haematobium were 6.1% and 3.7% using Kato Katz and urine filtration concentration testing techniques. The highest state prevalence was reported in Western Equatoria for both S. mansoni (14.7%) and S. haematobium (7.3%). The age of the participants varied from 4 to 18 years; of these, children 10 to 12 years old had the highest prevalence of S. mansoni (6.8%) and S. haematobium (3.7%). The prevalence of S. mansoni (7% male vs 5% female) and S. haematobium (3.6% male vs 3.1% female) were higher in males than females. The likelihood of the prevalence of S. mansoni in males was 1.42 (95% CI:1.23, 1.64) higher than in females, while for S. haematobium, 1.36 (95% CI:1.12, 1.65) higher than in females. The prevalence of S. mansoni and S. haematobium showed a statistically significant gender difference (P< 0.05). Conclusion: the study had provided evidence of the distribution of schistosomiasis in South Sudan for policy direction and recommended annual preventive chemotherapy with praziquantel in all endemic areas.

Diagnostic performance of two specific schistosoma japonicum immunological tests for screening schistosoma haematobium in school children in Zambia.

Dipstick Dye Immunoassay (DDIA) and Indirect Haemagglutination Assay (IHA), are two commercially available kits which have been widely used for screening Schistosoma japonicum in P.R. China. Whether they can be used for screening of Schistosoma haematobium are not clear. In order to evaluate the diagnostic efficiency of DDIA and IHA for screening Schistosoma haematobium, serum samples were collected from pupils in endemic areas in Zambia, Southern Africa, and tested by DDIA and IHA by single-blind manner. Meanwhile, the pupils were microscopically examined by infection with Schistosoma and soil-transmitted helminths, visually observed for parasite eggs. Of the enrolled 148 pupils, 61% tested positive for S. haematobium infection, while 31% and 36% of pupils were infected with hookworm and Ascaris respectively. Regarding the parasitological tests as reference standard, for the diagnosis of S. haematobium infection, IHA performed higher sensitivity (74%, 95% CI: 65%-83%) than that of DDIA (60%, 95%CI: 49%-70%). The sensitivities of IHA and DDIA are significant higher in 10-14 years old students than those of 7-9 years old group. The specificity of DDIA and IHA were 61% (95%CI: 49%-74%) and 72% (95%CI: 60%-84%), respectively. The co-infection with STHs decreased the specificity of DDIA but had no impact on that of IHA. Our study indicated that IHA has more potential as an alternative diagnostic tool for identifying schistosomiasis haematobium but need further improvement.

Performance of a real-time PCR approach for diagnosing Schistosoma haematobium infections of different intensity in urine samples from Zanzibar.

BACKGROUND: Efforts to control and eliminate schistosomiasis have accelerated over the past decade. As parasite burden, associated morbidity and egg excretion decrease, diagnosis with standard parasitological methods becomes harder. We assessed the robustness and performance of a real-time PCR (qPCR) approach in comparison with urine filtration microscopy and reagent strip testing for the diagnosis of Schistosoma haematobium infections of different intensities. METHODS: The robustness of DNA isolation and qPCR was validated in eight laboratories from Europe and Africa. Subsequently, 792 urine samples collected during cross-sectional surveys of the Zanzibar Elimination of Schistosomiasis Transmission (ZEST) project in 2012-2017 were examined with qPCR in 2018. Diagnostic sensitivity of the qPCR was calculated at different infection intensity categories, using urine filtration microscopy as reference test. Spearman's rank correlation between Ct-values and S. haematobium egg counts was assessed and Ct-value percentiles for infection intensity categories determined. RESULTS: S. haematobium Dra1 DNA-positive samples were identified correctly in all eight laboratories. Examination of urine samples from Zanzibar revealed Dra1 DNA in 26.8% (212/792) by qPCR, S. haematobium eggs in 13.3% (105/792) by urine filtration, and microhaematuria in 13.8% (109/792) by reagent strips. Sensitivity of the qPCR increased with augmenting egg counts: 80.6% (29/36) for counts between 1 and 4 eggs, 83.3% (15/18) for counts between 5 and 9 eggs, 100% (23/23) for counts between 10 and 49 eggs, and 96.4% (27/28) for counts of 50+ eggs. There was a significant negative correlation between Ct-values and egg counts (Spearman's rho = - 0.49, P < 0.001). Seventy-five percent of the Ct-values were ≥ 33 in the egg-negative category, < 31 in the light intensity category, and < 24 in the heavy intensity category. CONCLUSIONS: While the sensitivity of the qPCR was ~ 80% for very light intensity infections (egg counts < 10), in general, the Dra1 based qPCR assay detected twice as many S. haematobium infections compared with classical parasitological tests. The qPCR is hence a sensitive, urine-based approach for S. haematobium diagnosis that can be used for impact assessment of schistosomiasis elimination programmes, individual diagnosis, and in improved format also for verification and certification of elimination. TRIAL REGISTRATION: ISRCTN, ISRCTN48837681 . Registered 05 September 2012 - Retrospectively registered.

Elimination of schistosomiasis haematobia as a public health problem in five governorates in Upper Egypt.

The prevalence and intensity of Schistosoma haematobium infection was determined among schoolchildren living in five governorates in Upper Egypt. Between November 2016 and March 2017, urine samples were collected from 30,083 schoolchildren (6-16 years of age) from the governorates of Assiut (n = 7496; 6 districts), Bani Sweif (n = 4493; 7 districts), Fayoum (n = 4597; 6 districts), Menia (n = 7500; 9 districts) and Sohag (n = 5997; 11 districts). All samples were processed using urine filtration to detect and quantify S. haematobium eggs. The overall prevalence was 1.3% (95% Confidence Interval (CI) = 1.1%, 1.4%), but the prevalence varied considerably across districts in the studied governorates (from 0%, Fayoum to 13.4%, Sohag). The prevalence of heavy-intensity infections (≥50 egg/10 ml) varied from 0.05% (95% CI = 0.01-0.1) in Sohag to 0.3% (95% CI = 0.1-0.4) in Menia. No subject with heavy intensity of infection was detected in Fayoum and Bani Sweif governorates. Of the 39 studied districts 97.4% had prevalence of heavy intensity infection of <1%, indicating elimination of schistosomiasis haematobia as a public health problem in these districts. Of those studied 72.0% were male. Males were 2.9 times as likely to be infected (1.5% [95% CI: 1.4-1.7]) as females (0.5% [95% CI: 0.3-0.7]); χ2 = 51.2, p < 0.0001. Heavy intensity of infection was detected only in males. The prevalence of S. haematobium infection increased steadily with age, and the age group >15 years was 7 times as likely to be infected as the younger age group (6-<9; 0.8%); χ2 = 44.9, p < 0.0001. The national schistosomiasis control programme (NSCP) adopted a new elimination strategy by readjusting thresholds for MDA using praziquantel and targeting all transmission areas. The NSCP, after this major achievement of elimination of schistosomiasis haematobia as a public health problem, is now moving to interruption of its transmission.

Urogenital schistosomiasis elimination in Zanzibar: accuracy of urine filtration and haematuria reagent strips for diagnosing light intensity Schistosoma haematobium infections.

BACKGROUND: Urine filtration and microhaematuria reagent strips are basic standard diagnostic methods to detect urogenital schistosomiasis. We assessed their accuracy for the diagnosis of light intensity infections with Schistosoma haematobium as they occur in individuals living in Zanzibar, an area targeted for interruption of transmission. METHODS: Urine samples were collected from children and adults in surveys conducted annually in Zanzibar from 2013 through 2016 and examined with the urine filtration method to count S. haematobium eggs and with the reagent strip test (Hemastix) to detect microhaematuria as a proxy for infection. Ten percent of the urine filtration slides were read twice. Sensitivity was calculated for reagent strips, stratified by egg counts reflecting light intensity sub-groups, and kappa statistics for the agreement of urine filtration readings. RESULTS: Among the 39,207 and 18,155 urine samples examined from children and adults, respectively, 5.4% and 2.7% were S. haematobium egg-positive. A third (34.7%) and almost half (46.7%) of the egg-positive samples from children and adults, respectively, had ultra-low counts defined as 1-5 eggs per 10 ml urine. Sensitivity of the reagent strips increased significantly for each unit log10 egg count per 10 ml urine in children (odds ratio, OR: 4.7; 95% confidence interval, CI: 4.0-5.7; P < 0.0001) and adults (OR: 2.6; 95% CI: 1.9-3.7, P < 0.0001). Sensitivity for diagnosing ultra-light intensity infections was very low in children (50.1%; 95% CI: 46.5-53.8%) and adults (58.7%; 95% CI: 51.9-65.2%). Among the 4477 and 1566 urine filtration slides read twice from children and adults, most were correctly identified as negative or positive (kappa = 0.84 for children and kappa = 0.81 for adults). However, 294 and 75 slides had discrepant results and were positive in only one of the two readings. The majority of these discrepant slides (76.9% of children and 84.0% of adults) had counts of 1-5 eggs per 10 ml urine. CONCLUSIONS: We found that many individuals infected with S. haematobium in Zanzibar excrete > 5 eggs per 10 ml urine. These ultra-light infections impose a major challenge for accurate diagnosis. Next-generation diagnostic tools to be used in settings where interruption of transmission is the goal should reliably detect infections with ≤ 5 eggs per 10 ml urine. TRIAL REGISTRATION: ISRCTN, ISRCTN48837681 . Registered 05 September 2012 - Retrospectively registered.

A Prospective Blinded Evaluation of Urine-DNA Testing for Detection of Urothelial Bladder Carcinoma in Patients with Gross Hematuria.

Retrospective studies have provided proof of principle that bladder cancer can be detected by testing for the presence of tumor DNA in urine. We have conducted a prospective blinded study to determine whether a urine-based DNA test can replace flexible cystoscopy in the initial assessment of gross hematuria. A total of 475 consecutive patients underwent standard urological examination including flexible cystoscopy and computed tomography urography, and provided urine samples immediately before (n=461) and after (n=444) cystoscopy. Urine cells were collected using a filtration device and tested for eight DNA mutation and methylation biomarkers. Clinical evaluation identified 99 (20.8%) patients with urothelial bladder tumors. With this result as a reference and based on the analysis of all urine samples, the DNA test had a sensitivity of 97.0%, a specificity of 76.9%, a positive predictive value of 52.5%, and a negative predictive value of 99.0%. In three patients with a positive urine-DNA test without clinical evidence of cancer, a tumor was detected at repeat cystoscopy within 16 mo. Our results suggest that urine-DNA testing can be used to identify a large subgroup of patients with gross hematuria in whom cystoscopy is not required. PATIENT SUMMARY: We tested the possibility of using a urine-based DNA test to check for bladder cancer in patients with visible blood in the urine. Our results show that the test efficiently detects bladder cancer and therefore may be used to greatly reduce the number of patients who would need to undergo cystoscopy.

New diagnostic tools in schistosomiasis.

Schistosomiasis is a water-based parasitic disease that affects over 250 million people. Control efforts have long been in vain, which is one reason why schistosomiasis is considered a neglected tropical disease. However, since the new millennium, interventions against schistosomiasis are escalating. The initial impetus stems from a 2001 World Health Assembly resolution, urging member states to scale-up deworming of school-aged children with the anthelminthic drug praziquantel. Because praziquantel is safe, efficacious and inexpensive when delivered through the school platform, diagnosis before drug intervention was deemed unnecessary and not cost-effective. Hence, there was little interest in research and development of novel diagnostic tools. With the recent publication of the World Health Organization (WHO) Roadmap to overcome the impact of neglected tropical diseases in 2020, we have entered a new era. Elimination of schistosomiasis has become the buzzword and this has important ramifications for diagnostic tools. Indeed, measuring progress towards the WHO Roadmap and whether local elimination has been achieved requires highly accurate diagnostic assays. Here, we introduce target product profiles for diagnostic tools that are required for different stages of a schistosomiasis control programme. We provide an update of the latest developments in schistosomiasis diagnosis, including microscopic techniques, rapid diagnostic tests for antigen detection, polymerase chain reaction (PCR) assays and proxy markers for morbidity assessments. Particular emphasis is placed on challenges and solutions for new technologies to enter clinical practice.