RESUMO
BACKGROUND: The prevalence of intestinal parasites is known to be high among Amerindian populations; further, there are serious problems in the healthcare of these populations in Brazil. The Maxakali, located in the northeastern region of Minas Gerais, Brazil, is an indigenous group that still preserves many of its cultural aspects. This study aimed to compare the positivity rate of schistosomiasis and soil-transmitted helminths in this ethnic group in epidemiological surveys conducted in 1972 and 2014. METHODS: Stool parasitological examinations were performed by the Kato-Katz technique during both periods in this population. In 2014, the parasitological diagnosis was also realized with the TF-Test® technique. RESULTS: In 1972, 270 inhabitants were examined. The positivity rates were 67.4% for Schistosoma mansoni, 72.9% for hookworms, 43.7% for Ascaris lumbricoides, and 23.7% for Trichuris trichiura. In 2014, 545 individuals were examined, and the positivity rates obtained were 45.7% for S. mansoni, 22.8% for hookworms, 0.6% for A. lumbricoides, and 2.8% for T. trichiura. CONCLUSIONS: The comparison of the parasitological surveys conducted in 1972 and 2014, indicates that the indigenous Maxakali remained neglected by the health and indigenous protection authorities during these four decades. The infection rate observed in 2014 for schistosomiasis and hookworm remains high, considering the current epidemiological view of these diseases in the Brazilian population.
Assuntos
Esquistossomose , Humanos , Brasil/epidemiologia , Prevalência , Estudos Transversais , Fezes/parasitologia , Esquistossomose/epidemiologiaRESUMO
The laboratorial diagnosis of the intestinal schistosomiasis is always performed using Kato-Katz technique. However, this technique presents low sensitivity for diagnosis of individuals with low parasite burden, which constitutes the majority in low endemicity Brazilian locations for the disease. The objective of this study was developed and to validate a real-time PCR assay (qPCR) targeting 121 bp sequence to detect Schistosoma spp. DNA for the diagnosis of intestinal schistosomiasis and a sequence of the human ß-actin gene as internal control. Firstly, the qPCR was standardized and next it was evaluated for diagnosis and cure assessment of intestinal schistosomiasis in the resident individuals in Tabuas and Estreito de Miralta, two locations in Brazil endemic for intestinal schistosomiasis. The qPCR assay results were compared with those of the Kato-Katz (KK) test, examining 2 or 24 slides, Saline Gradient (SG) and "reference test" (24 KK slides + SG). The cure assessment was measured by these diagnostic techniques at 30, 90, and 180 days post-treatment. In Tabuas, the positivity rates obtained by the qPCR was 30.4% (45/148) and by "reference test" was of 31.0% (46/148), with no statistical difference (p = 0.91). The presumed cure rates at 30, 90, and 180 days post-treatment were 100, 94.4, and 78.4% by the analysis of 24 KK slides, 100, 94.4, and 78.4% by the SG, and 100, 83.3, and 62.1% by the qPCR assay. In Estreito de Miralta, the positivity obtained by qPCR was 18.3% (26/142) and with "reference test" was 24.6% (35/142), with no statistical difference (p = 0.20). The presumed cure rates were 93.3, 96.9, and 96.5% by the KK, 93.3, 96.9, and 100% by the SG, and 93.3, 93.9, and 96.5% by the qPCR at 30, 90, and 180 days post-treatment, respectively. This study showed that the diagnostic techniques presented different performance in the populations from the two districts (Tabuas and Estreito de Miralta) and reinforces the need of combining techniques to improve diagnosis accuracy, increasing the detection of individuals with low parasite burden. This combination of techniques consists an important strategy for controlling the disease transmission.
Assuntos
Anti-Helmínticos/uso terapêutico , DNA de Helmintos/análise , Fezes/parasitologia , Praziquantel/uso terapêutico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Criança , Pré-Escolar , Estudos Transversais , DNA de Helmintos/isolamento & purificação , Fezes/química , Feminino , Helmintos/genética , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Oligodesoxirribonucleotídeos/genética , Contagem de Ovos de Parasitas , Schistosoma mansoni/efeitos dos fármacos , Esquistossomose mansoni/tratamento farmacológico , Esquistossomose mansoni/parasitologia , Sensibilidade e Especificidade , Especificidade da Espécie , Resultado do Tratamento , Adulto JovemRESUMO
A prospective cohort study with rigorous searching for schistosomiasis cases was conducted among residents of Pedra Preta, Montes Claros, Minas Gerais State, Brazil, seven years after an intervention. Kato-Katz (KK), Saline Gradient, Miracidia Hatch and Polymerase Chain Reaction (PCR) were used as the diagnostic methods in 2008. In the period of 2013-2016, 175 patients remaining in the area were examined using the diagnostic methods Kato-Katz (24 slides, 1 g of feces) and Saline Gradient (2 procedures, 1 g of feces). Sixty-eight out of the 69 infected and treated individuals in 2008 tested negative. The percentage of new cases was 2.29% (4/175), and the 4 infected individuals presented low parasitic load [1, 6, 7 and 19 eggs per gram (EPG)]. All the participants answered epidemiological questionnaires on risky behavior. All residences had pit latrines and domiciliary water supply. The primary transmission focus (lake) was dry for several months. Malacological surveys showed a few non-infected specimens of Biomphalaria glabrata . A clear dominance of Biomphalaria straminea was observed. It can be inferred that a significant decrease in the disease transmission occurred after a single action through an intense search for infected and treated cases under the ecoepidemiological conditions of this area.
Assuntos
Biomphalaria/parasitologia , Praziquantel/administração & dosagem , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/tratamento farmacológico , Adulto , Animais , Estudos de Coortes , Feminino , Humanos , Masculino , Contagem de Ovos de Parasitas , Reação em Cadeia da Polimerase , Estudos Prospectivos , Esquistossomose mansoni/prevenção & controle , Esquistossomose mansoni/transmissão , Fatores SocioeconômicosRESUMO
The population of Brazil is currently characterised by many individuals harbouring low-intensity Schistosoma mansoni infections. The Kato-Katz technique is the diagnostic method recommended by the World Health Organization (WHO) to assess these infections, but this method is not sensitive enough in the context of low egg excretion. In this regard, potential alternatives are being employed to overcome the limits of the Kato-Katz technique. In the present review, we evaluated the performance of parasitological and immunological approaches adopted in Brazilian areas. Currently, the diagnostic choices involve a combination of strategies, including the utilisation of antibody methods to screen individuals and then subsequent confirmation of positive cases by intensive parasitological investigations.
Assuntos
Anticorpos Anti-Helmínticos/análise , Antígenos de Helmintos/análise , Técnicas de Laboratório Clínico/métodos , Fezes/parasitologia , Schistosoma mansoni , Esquistossomose mansoni/diagnóstico , Animais , Brasil/epidemiologia , Doenças Endêmicas , Humanos , Técnicas Imunoenzimáticas , Contagem de Ovos de Parasitas , Schistosoma mansoni/imunologia , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/epidemiologia , Sensibilidade e Especificidade , Índice de Gravidade de DoençaRESUMO
BACKGROUND: Despite decades of use of control programs, schistosomiasis remains a global public health problem. To further reduce prevalence and intensity of infection, or to achieve the goal of elimination in low-endemic areas, there needs to be better diagnostic tools to detect low-intensity infections in low-endemic areas in Brazil. The rationale for development of new diagnostic tools is that the current standard test Kato-Katz (KK) is not sensitive enough to detect low-intensity infections in low-endemic areas. In order to develop new diagnostic tools, we employed a proteomics approach to identify biomarkers associated with schistosome-specific immune responses in hopes of developing sensitive and specific new methods for immunodiagnosis. METHODS AND FINDINGS: Immunoproteomic analyses were performed on egg extracts of Schistosoma mansoni using pooled sera from infected or non-infected individuals from a low-endemic area of Brazil. Cross reactivity with other soil-transmitted helminths (STH) was determined using pooled sera from individuals uniquely infected with different helminths. Using this approach, we identified 23 targets recognized by schistosome acute and chronic sera samples. To identify immunoreactive targets that were likely glycan epitopes, we compared these targets to the immunoreactivity of spots treated with sodium metaperiodate oxidation of egg extract. This treatment yielded 12/23 spots maintaining immunoreactivity, suggesting that they were protein epitopes. From these 12 spots, 11 spots cross-reacted with sera from individuals infected with other STH and 10 spots cross-reacted with the negative control group. Spot number 5 was exclusively immunoreactive with sera from S. mansoni-infected groups in native and deglycosylated conditions and corresponds to Major Egg Antigen (MEA). We expressed MEA as a recombinant protein and showed a similar recognition pattern to that of the native protein via western blot. IgG-ELISA gave a sensitivity of 87.10% and specificity of 89.09% represented by area under the ROC curve of 0.95. IgG-ELISA performed better than the conventional KK (2 slides), identifying 56/64 cases harboring 1-10 eggs per gram of feces that were undiagnosed by KK parasitological technique. CONCLUSIONS: The serological proteome approach was able to identify a new diagnostic candidate. The recombinant egg antigen provided good performance in IgG-ELISA to detect individuals with extreme low-intensity infections (1 egg per gram of feces). Therefore, the IgG-ELISA using this newly identified recombinant MEA can be a useful tool combined with other techniques in low-endemic areas to determine the true prevalence of schistosome infection that is underestimated by the KK method. Further, to overcome the complexity of ELISA in the field, a second generation of antibody-based rapid diagnostic tests (RDT) can be developed.
Assuntos
Antígenos de Helmintos/sangue , Proteínas de Helminto/sangue , Proteoma/metabolismo , Schistosoma mansoni/imunologia , Esquistossomose mansoni/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Antígenos de Helmintos/imunologia , Biomarcadores/sangue , Brasil , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Feminino , Proteínas de Helminto/imunologia , Humanos , Imunoglobulina G/sangue , Lactente , Masculino , Pessoa de Meia-Idade , Óvulo/imunologia , Contagem de Ovos de Parasitas , Proteoma/imunologia , Proteômica , Proteínas Recombinantes/imunologia , Esquistossomose mansoni/sangue , Sensibilidade e Especificidade , Testes Sorológicos/métodosRESUMO
The population of Brazil is currently characterised by many individuals harbouring low-intensity Schistosoma mansoni infections. The Kato-Katz technique is the diagnostic method recommended by the World Health Organization (WHO) to assess these infections, but this method is not sensitive enough in the context of low egg excretion. In this regard, potential alternatives are being employed to overcome the limits of the Kato-Katz technique. In the present review, we evaluated the performance of parasitological and immunological approaches adopted in Brazilian areas. Currently, the diagnostic choices involve a combination of strategies, including the utilisation of antibody methods to screen individuals and then subsequent confirmation of positive cases by intensive parasitological investigations.
Assuntos
Humanos , Schistosoma mansoni , ImunoensaioRESUMO
Intestinal parasitic infections are a common health problem among Amerindian populations and schistosomiasis represents one of the most prevalent diseases in Maxakali people. The Kato-Katz is the diagnostic method recommended by WHO for epidemiological studies; however, one of the technique's limitations is the failure to detect parasites in individuals with low parasite load. The aim of this study was to establish the prevalence of Schistosoma mansoni in indigenous Maxakali villages, evaluating the TF-Test® performance for diagnosis compared to the Kato-Katz technique. Stool samples from 545 individuals were processed by the TF-Test® (1 sample) and Kato-Katz (1 slide). The positivity rate for S. mansoni by Kato-Katz was 45.7%. The rate by the TF-Test® was 33.2%, and 51.9% by the combined parasitological techniques. The amplitude of parasite load was 24 to 4,056 eggs per gram of feces (epg), with a geometric mean of 139 epg. The co-positivity, co-negativity, and accuracy values by TF-Test® in relation to Kato-Katz were 59.0%, 88.5%, and 75.0%, respectively. The agreement between these techniques was moderate (k=0.486) as determined by the kappa index. Thus, the results of this study demonstrated that the performance of Kato-Katz was superior (p <0.05) to that of TF-Test® in the detection of S. mansoni. The combination of TF-Test® and Kato-Katz resulted in an increased positivity rate of S. mansoni, demonstrating the high risk of infection to which indigenous populations are exposed and the importance of the implementation of control strategies in Maxakali villages.
Assuntos
Fezes/parasitologia , Indígenas Sul-Americanos/estatística & dados numéricos , Enteropatias Parasitárias/epidemiologia , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/epidemiologia , Adolescente , Adulto , Animais , Brasil/epidemiologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/parasitologia , Masculino , Pessoa de Meia-Idade , Contagem de Ovos de Parasitas , Carga Parasitária , Prevalência , Esquistossomose mansoni/diagnóstico , Sensibilidade e Especificidade , Adulto JovemRESUMO
OBJECTIVE: We developed a laboratorial platform to release a commercial platform used in the PCR-ELISA for the molecular diagnosis of schistosomiasis mansoni. On following, PCR-ELISA platform laboratorial was evaluated in 206 feces samples collected of individual living in a Brazilian low endemicity area. RESULTS: The PCR-ELISA laboratorial platform indicated a prevalence rate of 25.2%, which was higher than the Kato-Katz technique (18.4%) and lower than the commercial platform (30.1%). Considering Kato-Katz technique as the reference, there were 97.4% and 91.1% of relative sensitivity and specificity rates, respectively. The laboratorial platform presented good precision, performance diagnostic, and can be used in replacement to the commercial platform for diagnosis of schistosomiasis by PCR-ELISA.
Assuntos
Ensaio de Imunoadsorção Enzimática , Reação em Cadeia da Polimerase , Esquistossomose mansoni/diagnóstico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Brasil , Fezes/parasitologia , Feminino , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Contagem de Ovos de Parasitas , Prevalência , Schistosoma mansoni , Sensibilidade e Especificidade , Adulto JovemRESUMO
In order to effectively control and monitor schistosomiasis, new diagnostic methods are essential. Taking advantage of computational approaches provided by immunoinformatics and considering the availability of Schistosoma mansoni predicted proteome information, candidate antigens of schistosomiasis were selected and used in immunodiagnosis tests based on Enzime-linked Immunosorbent Assay (ELISA). The computational selection strategy was based on signal peptide prediction; low similarity to human proteins; B- and T-cell epitope prediction; location and expression in different parasite life stages within definitive host. Results of the above-mentioned analysis were parsed to extract meaningful biological information and loaded into a relational database developed to integrate them. In the end, seven proteins were selected and one B-cell linear epitope from each one of them was selected using B-cell epitope score and the presence of intrinsically disordered regions (IDRs). These predicted epitopes generated synthetic peptides that were used in ELISA assays to validate the rational strategy of in silico selection. ELISA was performed using sera from residents of areas of low endemicity for S. mansoni infection and also from healthy donors (HD), not living in an endemic area for schistosomiasis. Discrimination of negative (NEG) and positive (INF) individuals from endemic areas was performed using parasitological and molecular methods. All infected individuals were treated with praziquantel, and serum samples were obtained from them 30 and 180 days post-treatment (30DPT and 180DPT). Results revealed higher IgG levels in INF group than in HD and NEG groups when peptides 1, 3, 4, 5 and 7 were used. Moreover, using peptide 5, ELISA achieved the best performance, since it could discriminate between individuals living in an endemic area that were actively infected from those that were not (NEG, 30DPT, 180DPT groups). Our experimental results also indicate that the computational prediction approach developed is feasible for identifying promising candidates for the diagnosis of schistosomiasis and other diseases.
Assuntos
Epitopos/imunologia , Proteínas de Helminto/imunologia , Proteoma/imunologia , Schistosoma mansoni/imunologia , Esquistossomose/imunologia , Testes Sorológicos/métodos , Animais , Anti-Helmínticos/uso terapêutico , Estudos de Casos e Controles , Simulação por Computador , Epitopos/genética , Proteínas de Helminto/genética , Humanos , Imunoglobulina G/sangue , Praziquantel/uso terapêutico , Proteoma/genética , Schistosoma mansoni/genética , Esquistossomose/sangue , Esquistossomose/tratamento farmacológicoRESUMO
INTRODUCTION: Schistosomiasis, caused by Schistosoma mansoni, is a public health concern in Brazil. However, the most popular diagnostic method, the Kato-Katz technique, exhibits low sensitivity in low-endemicity areas. We aimed to compare the performance of an immunological assay, the point-of-care circulating cathodic antigen (POC-CCA®) test, with that of two parasitological techniques in a low-endemicity population. METHODS: Our study included 141 individuals living in Estreito de Miralta, Minas Gerais, Brazil. Fecal samples were obtained from all participants and analyzed for schistosomiasis using two parasitological techniques: the Kato-Katz technique and the saline gradient technique. Additionally, POC-CCA® strips were utilized for testing urine samples. The results obtained by the different techniques were compared. RESULTS: Analysis of two or 24 slides using the Kato-Katz technique resulted in a positivity rate of 10.6% (15/141) or 19.1% (27/141), respectively. The saline gradient technique yielded a positivity rate of 17.0% (24/141). The prevalence according to both parasitological techniques was 24.1% (34/141). The POC-CCA® test yielded a positivity rate of 22.7% (32/141); however, the positivity rate was merely 2.1% if trace results were considered negative. The agreements observed between POC-CCA® and the parasitological techniques were good (Kappa indexes > 0.64). The POC-CCA® test was more sensitive than the two-slide Kato-Katz technique (p < 0.05) in detecting cases of S. mansoni infection when trace results were considered positive. CONCLUSIONS: These findings reinforce the importance of using multiple diagnostic techniques in low-endemicity areas for effective control of disease.