Abstract. Substantial uncertainties surround the sensitivities and specificities of diagnostic techniques for urinary schistosomiasis. We used latent class (LC) modeling to address this problem. In this study, 220 adults in three villages northwest of Accra, Ghana were examined using five Schistosoma haematobium diagnostic measures: microscopic examination of urine for detection of S. haematobium eggs, dipsticks for detection of hematuria, tests for circulating antigens, antibody tests, and ultrasound scans of the urinary system. Testing of the LC model indicated non-invariance of the performance of the diagnostic tests across different age groups, and measurement invariance held for males and females and for the three villages. We therefore recommend the use of LC models for comparison between and the identification of the most accurate schistosomiasis diagnostic tests. Furthermore, microscopy and hematuria dipsticks were indicated through these models as the most appropriate techniques for detection of S. haematobium infection.

Long-term infection with urinary schistosomiasis has been associated with development of bladder cancer. However, bladder cancer is difficult to diagnose without invasive measures such as cystoscopy, thus there is little information on the epidemiological extent of the problem. Studies have been either case-control studies or case examinations in different geographical areas, estimating a schistosome-associated bladder cancer incidence of 3-4 cases per 100,000. We have used three indicators to examine the potential bladder cancer problem in an adult rural population in Ghana endemic for urinary schistosomiasis: (i) parasitological positivity; (ii) age prevalence of bladder damage from ultrasound scans; and (iii) detection of biomarkers associated with the presence of bladder cancer. Biomarkers were BLCA-4 test (urine) and nuclear morphometry or quantitative nuclear grading (QNG) of epithelial cells (urine sediment), which quantifies DNA ploidy status and nuclear morphometric descriptors, both of which can detect the presence of bladder cancer. Our data show an increasing association between age, severe bladder abnormalities and the occurrence of these biomarkers. Sixty-two of 73 cytopathology Papanicolaou-stained smears were seen to have squamous metaplasia. Although further investigations are needed, we suggest that schistosome-associated bladder cancer is an important public health concern in areas where Schistosoma haematobium is prevalent.

Differential diagnosis of Schistosoma mansoni and S. haematobium, which often occur sympatrically in Africa, requires both urine and stool and the procedures are low in sensitivity. The standard diagnostic tests, such as Kato-Katz (KK) for S. mansoni eggs and presence of haematuria for S. haematobium both lack sensitivity, produce false-negative results and show reduced accuracy with decreasing intensity of infection. The need for a single diagnostic test with high sensitivity and specificity for both parasites is important as many African countries are implementing Mass Drug Administration (MDA) following recommendations of the World Health Organization (WHO). Eighty-six samples of urine sediment obtained by filtration were collected from a group of 5–23 years old people from an endemic area of southern Ghana. DNA was extracted from the urine sediment on filter paper from which a species-specific repeat fragment was amplified by polymerase chain reaction (PCR) with specific primers for S. mansoni and for S. haematobium. Additionally, all participants were tested by KK (stool) and dipstick for haematuria. Diagnostic parameters for all three tests were analyzed statistically. Amplification of species-specific DNA by PCR showed much higher sensitivity (99%–100%) and specificity (100%) compared to KK and haematuria (sensitivity: 76% and 30% respectively) for both schistosome species. The same pattern was observed when the data were stratified for age group and sex specific analysis. In addition PCR amplification detected DNA from 11 individuals infected with both parasites who were negative by KK and haematuria. This approach of detecting parasite specific DNA from either or both species in a single urine specimen is a practical advantage that avoids the need for two specimens and is more effective than standard tests including those based on serology. This promises to improve the effectiveness of surveillance of MDA control programs of schistosomiasis.

Purpose Schistosoma haematobium is associated with chronic bladder damage and may subsequently induce bladder cancer in humans, thus posing a serious threat where the parasite is endemic. Here we evaluated aberrant promoter DNA methylation as a potential biomarker to detect severe bladder damage that is associated with schistosomiasis by analyzing urine specimens.Materials and MethodsA quantitative methylation-specific PCR (QMSP) assay was used to examine the methylation status of seven genes (RASSF1A, RARβ2, RUNX3, TIMP3, MGMT, P16, ARF) in 57 urine samples obtained from volunteers that include infected and uninfected by S. haematobium from an endemic region. The Fishers Exact Test and Logistic Regression analysis were used to evaluate the methylation status with bladder damage (as assessed by ultrasound examination) in subjects with S. haematobium infection.Results RASSF1A and TIMP3 were significant to predict severe bladder damage both in univariate (p = 0.015 and 0.023 respectively) and in multivariate (p = 0.022 and 0.032 respectively) logistic regression analysis. Area under the receiver operator characteristic curves (AUC-ROC) for RASSF1A and TIMP3 to predict severe bladder damage were 67.84% and 63.73% respectively. The combined model, which used both RASSF1A and TIMP3 promoter methylation, resulted in significant increase in AUC-ROC compared to that of TIMP3 (77.55% vs. 63.73%.29; p = 0.023).ConclusionsIn this pilot study, we showed that aberrant promoter methylation of RASSF1A and TIMP3 are present in urine sediments of patients with severe bladder damage associated with S. haematobium infection and that may be used to develop non-invasive biomarker of S. haematobium exposure and early molecular risk assessmentof neoplastic transformation.

The association between chronic infection with Schistosoma haematobium and squamous cell carcinoma of the bladder is well known but there are few epidemiological data available to assess the extent of this cancer in schistosome-endemic areas. Invasive surgical procedures are not practical for epidemiological appraisal, therefore this important health matter is unresolved. This review examines recent work done to identify and detect biomarkers that can be found in voided urine and therefore obtained without invasive procedures. A variety of products of cell cycle kinetics, nuclear activity enzymes and even nuclear morphometry have been studied in urine specimens and these in concert may be sufficient to indicate the likelihood of the presence of bladder cancer or some predictive pre-cancerous state. Although these techniques require a sophisticated central laboratory for analysis, specimens can easily be collected in the field and brought to the centre, which could serve regional programmes. We suggest that early diagnosis of pre-cancerous conditions associated with urinary schistosomiasis, if appropriately treated, could save countless lives.