Eduardo Brandão

Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims to eliminate this disease by the year 2020. However, the development of more specific and sensitive tests is important for the success of the GPELF. The present study aimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosis of filariasis in serum and urine. Twenty paired biological urine and serum samples from individuals already known to be positive for Wuchereria bancrofti were collected during the day. Conventional PCR and semi-nested PCR assays were optimised. The detection limit of the technique for purified W. bancrofti DNA extracted from adult worms was 10 fg for the internal systems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primers was confirmed experimentally by amplification of 1 ng of purified genomic DNA from other species of parasites. Evaluation of the paired urine and serum samples by the semi-nested PCR technique indicated only two of the 20 tested individuals were positive, whereas the simple internal PCR system (WbF/Wb2), which has highly promising performance, revealed that all the patients were positive using both samples. This study successfully demonstrated the possibility of using the PCR technique on urine for the diagnosis of W. bancrofti infection.

Epidemiological Assessment of Neglected Diseases in Children: Lymphatic Filariasis and Soil-Transmitted Helminthiasis

OBJECTIVE To report the prevalence of lymphatic filariasis and intestinal parasitic infections in school-aged children living in a filariasis endemic area and discuss about the therapeutic regimen adopted in Brazil for the large-scale treatment of filariasis. METHODS A cross-sectional study including 508 students aged 5-18 years old, enrolled in public schools within the city of Olinda, Pernambuco. The presence of intestinal parasites was analyzed using the Hoffman, Pons and Janer method on 3 stool samples. The diagnosis of filarial infection was performed using the rapid immunochromatographic technique (ICT) for the antigen, and the polycarbonate membrane filtration for the presence of microfilariae. Descriptive statistics of the data was performed using EpiInfo version 7. RESULTS The prevalence of filariasis was 13.8% by ICT and 1.2% by microfilaraemia, while intestinal parasites were detected in 64.2% of cases. Concurrent diagnosis of filariasis and intestinal parasites was 9.4%, while 31.5% of students were parasite-free. Among individuals with intestinal parasites, 55% had one parasite and 45% had more than one parasite. Geohelminths occurred in 72.5% of the parasited individuals. In the group with filarial infection the prevalence of soil-transmitted helminthiasis was 54.5%. CONCLUSIONS The simultaneous diagnosis of filariasis and intestinal parasites as well as the high frequency of geohelminths justify the need to reevaluate the treatment strategy used in the Brazilian filariasis large-scale treatment program.Objective To report the prevalence of lymphatic filariasis and intestinal parasitic infections in school-aged children living in a filariasis endemic area and discuss about the therapeutic regimen adopted in Brazil for the large-scale treatment of filariasis.

Evaluation of diagnostic tests for Wuchereria bancrofti infection in Brazilian schoolchildren

INTRODUCTION Since the launch of the Global Programme to Eliminate Lymphatic Filariasis, more than 70% of the endemic countries have implemented mass drug administration (MDA) to interrupt disease transmission. The monitoring of filarial infection in sentinel populations, particularly schoolchildren, is recommended to assess the impact of MDA. A key issue is choosing the appropriate tools for these initial assessments (to define the best intervention) and for monitoring transmission. METHODS This study compared the pre-MDA performance of five diagnostic methods, namely, thick film test, Knotts technique, filtration, Og4C3-ELISA, and the AD12-ICT card test, in schoolchildren from Brazil. Venous and capillary blood samples were collected between 11 pm and 1 am. The microfilarial loads were analyzed with a negative binomial regression, and the prevalence and associated 95% confidence intervals were estimated for all methods. The accuracies of the AD12-ICT card and Og4C3-ELISA tests were assessed against the combination of parasitological test results. RESULTS A total of 805 schoolchildren were examined. The overall and stratified prevalence by age group and gender detected by Og4C3-ELISA and AD12-ICT were markedly higher than the prevalence estimated by the parasitological methods. The sensitivity of the AD12-ICT card and Og4C3-ELISA tests was approximately 100%, and the positive likelihood ratios were above 6. The specificity of the Og4C3-ELISA was higher than that of the AD12-ICT at different prevalence levels. CONCLUSIONS The ICT card test should be the recommended tool for monitoring school-age populations living in areas with ongoing or completed MDA.

Original articleEpidemiological assessment of neglected diseases in children: lymphatic filariasis and soil-transmitted helminthiasisPrevalência de doenças negligenciadas em escolares: filariose linfática e parasitoses intestinais☆

OBJECTIVE To report the prevalence of lymphatic filariasis and intestinal parasitic infections in school-aged children living in a filariasis endemic area and discuss about the therapeutic regimen adopted in Brazil for the large-scale treatment of filariasis. METHODS A cross-sectional study including 508 students aged 5-18 years old, enrolled in public schools within the city of Olinda, Pernambuco. The presence of intestinal parasites was analyzed using the Hoffman, Pons and Janer method on 3 stool samples. The diagnosis of filarial infection was performed using the rapid immunochromatographic technique (ICT) for the antigen, and the polycarbonate membrane filtration for the presence of microfilariae. Descriptive statistics of the data was performed using EpiInfo version 7. RESULTS The prevalence of filariasis was 13.8% by ICT and 1.2% by microfilaraemia, while intestinal parasites were detected in 64.2% of cases. Concurrent diagnosis of filariasis and intestinal parasites was 9.4%, while 31.5% of students were parasite-free. Among individuals with intestinal parasites, 55% had one parasite and 45% had more than one parasite. Geohelminths occurred in 72.5% of the parasited individuals. In the group with filarial infection the prevalence of soil-transmitted helminthiasis was 54.5%. CONCLUSIONS The simultaneous diagnosis of filariasis and intestinal parasites as well as the high frequency of geohelminths justify the need to reevaluate the treatment strategy used in the Brazilian filariasis large-scale treatment program.Objective To report the prevalence of lymphatic filariasis and intestinal parasitic infections in school-aged children living in a filariasis endemic area and discuss about the therapeutic regimen adopted in Brazil for the large-scale treatment of filariasis.

Lymphatic filariasis: Surveillance action among immigrants from endemic areas, Acre State, Brazilian Amazon

Abstract Objective To investigate the positivity of Wuchereria bancrofti ( W. bancrofti ) in immigrants who entered the country through Rio Branco, reducing the risk of introduction of parasites into new areas and endemic areas of the past. Methods It was realized a descriptive study. The AD12-ICT card test was applied on all immigrants living temporarily in the Chacara Alliance shelter, located in the metropolitan area of Rio Branco-AC, Brazil. For the positive patients, 10 mL of venous blood was collected between 11:00 pm and 1:00 am. About 4 mL of venous blood was collected to detect the presence of microfilariae in circulation in the tube using ethylene diamine tetraacetic acid and 6 mL of venous blood was collected to obtain blood serum for the Og4C3-ELISA, antibody Bm-14 and DNA- W. bancrofti tests. Results The present study evaluated 415 individuals in September 2014 by circulating filarial antigen for W. bancrofti using the AD12-ICT card test. A total of 15/415 (3.61%) positive cases were found, all from Haiti. Night blood collection and serum were performed on 1/14 for confirmation the infection of W. bancrofti , which presented 34 microfilariae/mL, antigen, antibodies and PCR positives. Conclusions This surveillance action reveals, in a pioneering and unequivocal manner, that Brazil is an influx of immigrants carrying lymphatic filariasis and there is an urgent need to step up surveillance at the main entry point for immigrants. Active surveillance may prevent the reintroduction of lymphatic filariasis in areas under control, or prevent its introduction into other states of Brazil.

Burden of lymphatic filariasis morbidity in an area of low endemicity in Brazil.

The Global Programme to Eliminate Lymphatic Filariasis has two main components: interrupting transmission of lymphatic filariasis (LF) and managing morbidity and preventing disability. However, interventions to prevent and manage LF-related disabilities in endemic communities have been of limited extent. The aim of this study was to describe the prevalence of morbidity and its correlation with filarial infection, thereby filling a gap that existed regarding the data on morbidity in Brazil. Presence of Wuchereria bancrofti microfilaria was investigated using the thick smear technique. Information on parasitosis-related clinical manifestations was obtained using a questionnaire applied by community health agents with previous training and capacitation to know about and identify the disease. To analyze correlations, Pearsons correlation coefficient was used with the corresponding statistical significance test. 23,673 individuals were investigated: 323 presented microfilaremia (1.36%) and 741 (3.13%) had clinical complaints that were attributable to LF. Acute dermatolymphangioadenitis (ADLA) was the most prevalent condition (2.2%). Lymphedema, ADLA and chyluria were more commonly reported among female patients. There were positive associations between all the clinical complaints reported and filarial infection. Hydrocele presented the most strongly positive association (r=0.699; p<0.001). The present study showed that there is an association between clinical condition reported and the rate of infection among people living in an area of low endemicity for LF. It contributes data that might provide support for healthcare systems and thus optimize disease management, through incorporating surveillance measures directed towards preventing disability and reducing the psychosocial and economic impact of the disease on poor populations living in areas endemic for LF.

Using kernel density estimates to investigate lymphatic filariasis in northeast Brazil

Abstract After more than 10 years of the Global Program to Eliminate Lymphatic Filariasis (GPELF) in Brazil, advances have been seen, but the endemic disease persists as a public health problem. The aim of this study was to describe the spatial distribution of lymphatic filariasis in the municipality of Jaboatão dos Guararapes, Pernambuco, Brazil. An epidemiological survey was conducted in the municipality, and positive filariasis cases identified in this survey were georeferenced in point form, using the GPS. A kernel intensity estimator was applied to identify clusters with greater intensity of cases. We examined 23 673 individuals and 323 individuals with microfilaremia were identified, representing a mean prevalence rate of 1·4%. Around 88% of the districts surveyed presented cases of filarial infection, with prevalences of 0–5·6%. The male population was more affected by the infection, with 63·8% of the cases (P<0·005). Positive cases were found in all age groups examined. The kernel intensity estimator identified the areas of greatest intensity and least intensity of filarial infection cases. The case distribution was heterogeneous across the municipality. The kernel estimator identified spatial clusters of cases, thus indicating locations with greater intensity of transmission. The main advantage of this type of analysis lies in its ability to rapidly and easily show areas with the highest concentration of cases, thereby contributing towards planning, monitoring, and surveillance of filariasis elimination actions. Incorporation of geoprocessing and spatial analysis techniques constitutes an important tool for use within the GPELF.

Effectiveness of annual single doses of diethylcarbamazine citrate among bancroftian filariasis infected individuals in an endemic area under mass drug administration in Brazil

ABSTRACT The Global Program to Eliminate Lymphatic Filariasis has achieved extraordinary success in reducing transmission and preventing morbidity through mass drug administration (MDA) to the population at-risk. Brazil is the only currently using diethylcarbamazine citrate (DEC) alone for MDA, so an assessment of its effectiveness is needed. We report the trends of filarial markers in a cohort of 175 individuals infected with Wuchereria bancrofti in areas that underwent MDA in the city of Olinda, Northeastern Brazil. The prospective study was conducted between 2007 and 2012 (corresponding to five annual MDA rounds). The quantification of microfilaraemia (QMFF) was assessed by filtration. Circulating filarial antigen (CFA) was detected through immunochromatographic point-of-care test (POCT-ICT) and Og4C3-ELISA whereas antifilarial antibody titres (IgG4) were assessed through Bm14 assay. The CFA and IgG4 titres were measured by Optical Density (OD). The main characteristics at baseline, MDA coverage and the trend of filarial infection markers during follow up were described. The trend of filarial markers in relation to time (years of MDA), sex and age were analysed through Generalized Estimating Equations (GEE) models. The models demonstrated a significant decrease in all markers during MDA. The probability of remaining positive by QMFF and POCT-ICT diminished 70% and 46%, respectively, after each MDA round. There was a significant annual drop in CFA (−0.290 OD) and IgG4 antibodies titres (−0.303 OD). This study provides evidence that MDA with DEC alone can be effective in the elimination of LF in Brazil.

Características clínico-epidemiológicas de la estrongiloidiasis en pacientes portadores de co-morbilidades

The strongyloidiasis is a parasitic disease that poses as a serious public health problem, mainly in tropical and subtropical countries. Over the years, some conditions, such as advances in corticosteroid treatment and immunosuppressive diseases, have improved not only the increase in cases of strongyloidiasis, but also the emergence of severe forms of the disease and / or deaths. For these reasons, the objective of this study is to make a critical analysis of the occurrence of strongyloidiasis in patients with comorbidities, describing clinical and epidemiological characteristics associated with these diseases that can highlight the importance of monitoring this parasitosis in most susceptible groups.

Screening and evaluation of lymphatic filariasis in immigrants from endemic countries residing in a focus where it is considered eliminated in the Southern Region of Brazil: A risk of reemergence?

Lymphatic filariasis (LF) has been targeted by the World Health Organization for elimination by the year 2020. However, migration of infected individuals from areas where LF is endemic to areas considered non-endemic or foci for the control and elimination may jeopardize the achievement of this goal. The aim of the present study was to evaluate the occurrence of filarial infection by way of circulating filarial antigen (CFA) circulation using the point of care AD12-immunochromatography card (POC-ICT) among immigrants from Haiti residing in Chapecó, Santa Catarina, between May and October 2015. Of the 420 subjects examined, 77.4% were male, aged 19-54 years. Ten (2.38%) were POC-ICT positive. Of this total, one was not found. Two individuals were negative for Og4C3-ELISA and DNA/Wb-PCR in all biological samples, but positive for the anti-filarial antibody Bm14 and only one showed microfilaremia (1mf/mL). These findings point to the importance of the Brazilian surveillance action to reduce the possibility of reintroduction of LF in Chapecó, Santa Catarina, by infected immigrants, and to guarantee the success of the National LF Elimination Plan.