N.K. Gunawardena

The use of whole blood absorbed on filter paper to detect Wuchereria bancrofti circulating antigen

The Og4C3 enzyme-linked immunosorbent assay (ELISA) to detect circulating Wuchereria bancrofti antigen uses 50 microL of serum. In this study, a whole blood sample absorbed on filter paper was tested as a substitute for serum. Serum samples were obtained from 60 Sri Lankan subjects by venepuncture and finger-prick blood samples from the same individuals were directly absorbed on filter paper. Og4C3 ELISAs using serum and filter paper blood were compared. Despite the fact that the estimated amount of serum available for the ELISA with filter paper blood was only one-fifth of that available when serum was used, the 2 ELISAs gave almost identical results. Of the 39 positive serum samples, 38 were detected using filter paper blood. Employing the ELISA using filter paper blood, 619 people in Matara, Sri Lanka, were examined for antigenaemia. The positivity rate was 22.5%, 3.1 times higher than the rate of microfilaraemia detected by examination of 60 microL blood films.

Epidemiology of bancroftian filariasis in three suburban areas of Matara, Sri Lanka

The epidemiological parameters of bancroftian filariasis were investigated in three suburbs of Matara, within the south-western coastal belt of Sri Lanka where the disease is endemic. The overall prevalence of microfilaraemia and the geometric mean density of the microfilaraemias observed were 4.4% and 20.6 microfilariae/60 µl fingerprick blood, respectively. Prevalence was significantly lower in the female subjects than in the male, and in males aged <20 years than in older males. Overall, 9.5% of the subjects had the clinical manifestations of bancroftian filariasis (6.4% had filarial fever, 3.0% had elephantiasis and/or oedema, and 6.2% had hydrocele). The prevalence of elephantiasis/oedema was generally higher among the female subjects (4.2%) than among the male (1.4%), and an age-prevalence plot for this manifestation showed a linear increase in prevalence after the age of 40 years. Hydrocele also became commoner with increasing age, but this increase in prevalence began at the lower age of 20 years. More than 60% of the cases of elephantiasis/oedema but only 26.3% of the subjects found to have hydroceles experienced filarial fever attacks. The cases of fever and elephantiasis/oedema (but not those of microfilaraemia or hydrocele) were aggregated within households. However, the children whose mothers were microfilaraemic were much more likely to be microfilaraemic themselves (8.7%) than the children who had amicrofilaraemic mothers (2.8%), microfilaraemic fathers (0.0%) or amicrofilaraemic fathers (2.7%). The results of entomological surveys indicated that transmission of Wuchereria bancrofti occurred throughout the year in the study community.

Human infection with Wuchereria bancrofti in Matara, Sri Lanka : the use, in parallel, of an ELISA to detect filaria-specific IgG4 in urine and of ICT card tests to detect filarial antigen in whole blood

Abstract The ICT card test to detect circulating filarial antigen and an ELISA that detects filaria-specific urinary IgG4 were each used to screen 473 subjects from a community in Sri Lanka where Wuchereria bancrofti is endemic. When the ICT test was used as the gold standard, the ELISA was found to have a sensitivity of 91.2%. However, far more of the subjects were found ELISA-positive than ICT-positive (76.5% v. 31.1%). The youngest children studied (aged 1-10 years) were similar to the adult subjects in terms of the prevalence of antigenaemia (33.8%) and the prevalence (72.1%) and concentration of filaria-specific IgG4 in their urine. Therefore, especially as urine samples are easier, less painful and safer to collect than blood samples, the ELISA may be particularly useful to screen very young and school-age children, to estimate current levels of transmission in a particular area.

Mapping the Risk of Snakebite in Sri Lanka - A National Survey with Geospatial Analysis.

Background There is a paucity of robust epidemiological data on snakebite, and data available from hospitals and localized or time-limited surveys have major limitations. No study has investigated the incidence of snakebite across a whole country. We undertook a community-based national survey and model based geostatistics to determine incidence, envenoming, mortality and geographical pattern of snakebite in Sri Lanka. Methodology/Principal Findings The survey was designed to sample a population distributed equally among the nine provinces of the country. The number of data collection clusters was divided among districts in proportion to their population. Within districts clusters were randomly selected. Population based incidence of snakebite and significant envenoming were estimated. Model-based geostatistics was used to develop snakebite risk maps for Sri Lanka. 1118 of the total of 14022 GN divisions with a population of 165665 (0.8%of the country’s population) were surveyed. The crude overall community incidence of snakebite, envenoming and mortality were 398 (95% CI: 356–441), 151 (130–173) and 2.3 (0.2–4.4) per 100000 population, respectively. Risk maps showed wide variation in incidence within the country, and snakebite hotspots and cold spots were determined by considering the probability of exceeding the national incidence. Conclusions/Significance This study provides community based incidence rates of snakebite and envenoming for Sri Lanka. The within-country spatial variation of bites can inform healthcare decision making and highlights the limitations associated with estimates of incidence from hospital data or localized surveys. Our methods are replicable, and these models can be adapted to other geographic regions after re-estimating spatial covariance parameters for the particular region.

Integrated School-Based Surveillance for Soil-Transmitted Helminth Infections and Lymphatic Filariasis in Gampaha District, Sri Lanka

We explored the practicality of integrating surveillance for soil-transmitted helminthiasis (STH, assessed by Kato-Katz) with transmission assessment surveys for lymphatic filariasis (LF) in two evaluation units (EUs) in Gampaha district, Sri Lanka (population 2.3 million). The surveys were performed 6 years after five annual rounds of mass drug administration with diethylcarbamazine and albendazole. Each transmission assessment survey tested children (N = 1,462 inland EU; 1,642 coastal EU) sampled from 30 primary schools. Low filarial antigenemia rates (0% and 0.1% for the inland and coastal EUs) suggest that LF transmission is very low in this district. The STH rates and stool sample participation rates were 0.8% and 61% (inland) and 2.8% and 58% (coastal). Most STH detected were low or moderate intensity Trichuris trichiura infections. The added cost of including STH testing was ∼

Impact of helminth infection on childhood allergic diseases in an area in transition from high to low infection burden

5,000 per EU. These results suggest that it is feasible to integrate school-based surveillance for STH and LF.

Isothermal Diagnostic Assays for monitoring single nucleotide polymorphisms in Necator americanus associated with Benzimidazole drug resistance

Background The effect of helminth infections on allergic diseases is still inconclusive. Furthermore, the effect of helminth infections on childhood allergic diseases in a tropical area where prevalence of helminth infections has undergone dramatic changes is not well documented. Objective To investigate the relationship between allergic diseases and helminth infection in a cohort of schoolchildren in an area that has undergone dramatic changes in intensity of helminth infections. Methods Children attending grade 5 were recruited from 17 schools in Western Province of Sri Lanka. They were assessed for allergic diseases using the International Study of Asthma and Allergies in Childhood questionnaire. Their serum total IgE (tIgE) and allergen-specific IgE (sIgE) for five common aeroallergens were measured by ImmunoCAP® method and stools were examined for the presence of helminth infections. Results A total of 640 children (mean age 10 years) were recruited to the study. Of them, 33.7% had evidence of allergic disease and 15.5% had helminth infections. Majority of infections (68.9%) were of low intensity. A significant relationship between allergic disease and helminth infections was not observed, however, a trend toward protective role of helminth infections against allergic diseases was noted. Multivariate analysis showed helminth infections to be an independent predictor of high tIgE levels whereas allergic disease was not. Allergic sensitization (atopy) was a significant risk factor for allergic disease only among non-infected children (odds ratio 3.025, p = 0.022) but not in infected children. The ratio of sIgE to tIgE was higher in non-infected children. Conclusion Though not significant, a reduced risk of allergy in helminth-infected children was observed in this population. A Decrease in intensity of helminth infections may have contributed to the reduced capacity of immune-modulation by helminths in this paediatric population.

Epidemiology of irritable bowel syndrome in children and adolescents in Asia.

Background Soil-transmitted helminths (STHs) are the most prevalent intestinal helminths of humans, and a major cause of morbidity in tropical and subtropical countries. The benzimidazole (BZ) drugs albendazole (ABZ) and mebendazole (MBZ) are used for treatment of human STH infections and this use is increasing dramatically with massive drug donations. Frequent and prolonged use of these drugs could lead to the emergence of anthelmintic resistance as has occurred in nematodes of livestock. Previous molecular assays for putative resistance mutations have been based mainly on PCR amplification and sequencing. However, these techniques are complicated and time consuming and not suitable for resource-constrained situations. A simple, rapid and sensitive genotyping method is required to monitor for possible developing resistance to BZ drugs. Methods To address this problem, single nucleotide polymorphism (SNP) detection assays were developed based on the Smart amplification method (SmartAmp2) to target codons 167, 198, and 200 in the β-tubulin isotype 1 gene for the hookworm Necator americanus. Findings Diagnostic assays were developed and applied to analyze hookworm samples by both SmartAmp2 and conventional sequencing methods and the results showed high concordance. Additionally, fecal samples spiked with N. americanus larvae were assessed and the results showed that the Aac polymerase used has high tolerance to inhibitors in fecal samples. Conclusion The N. americanus SmartAmp2 SNP detection assay is a new genotyping tool that is rapid, sensitive, highly specific and efficient with the potential to be used as a field tool for monitoring SNPs associated with BZ resistance. However, further validation on large numbers of field samples is required.

Prevalence and intensity of Wuchereria bancrofti antigenaemia in Sri Lanka by Og4C3 ELISA using filter paper-absorbed whole blood

Objectives: Several cross-sectional surveys have been conducted to study the prevalence of irritable bowel syndrome (IBS) in children. The aim of the present study was to conduct a systematic review and a meta-analysis of published literature to estimate the prevalence of IBS among Asian children. Method: A computer-assisted search of MEDLINE, EMBASE, PsycINFO, and regional databases of Asia was carried out. Selected articles were reviewed in depth and data were extracted. Pooled prevalence, sex differences, and 95% confidence intervals were calculated. Heterogeneity of the studies was assessed using the I2 test. Results: A total of 16 cross-sectional studies that reported prevalence of IBS (in children and adolescents) and qualified to be included were taken into the final analysis containing 38,076 patients. Selected studies are from China, Korea, Japan, Iran, Sri Lanka, and Saudi Arabia. Studies showed a marked heterogeneity with I2 of 98.59 (P < 0.0001). Prevalence of IBS ranges from 2.8% to 25.7%, with a pooled prevalence of 12.41% (95% confidence interval 9.87–14.95). Prevalence risk ratio of girl:boy is 1.39. Prevalence of subtypes is diverse and varies between studies. Conclusions: The published data indicate that IBS is a significant problem among Asian children and adolescents. Female sex predisposes children and adolescents to develop IBS.

Prevalence of enterobiasis among primary school children in Ragama, Sri Lanka.

In Sri Lanka 2741 people from Matara, an endemic area for Wuchereria bancrofti, were examined in 1996/97 for microfilariae by 60-microL blood smear and for circulating filarial antigens by Og4C3 ELISA using filter paper-absorbed whole blood. The overall prevalence of microfilaraemia was 3.4%, and that of antigenaemia 14.4%. The prevalence of antigen-positive and microfilaria-negative people was 11.3%. Analysed by age-group, antigenaemia prevalence was similar in all groups, and the average number of antigen units was already very high in the age-group < 10 years, indicating that the infection started in early childhood. Among those who were antigen positive, the microfilaria prevalence was lower in females than in males. Diethylcarbamazine treatment eliminated microfilariae in 78% of the positives. However, 17 months after the treatment, antigenaemia was still positive in 76% of those who were parasitologically cured.