Evaluation of immunological, parasitological and molecular methods for the diagnosis of Schistosoma mansoni infection before and after chemotherapy treatment with praziquantel in experimentally infected Nectomys squamipes.

In low endemicity areas of schistosomiasis, the recommended diagnostic method of coprological examination results in an underestimation of infection cases. Alternative diagnostic methods have been developed, such as immunodiagnostic and molecular techniques. In this study we evaluated three methods used in the diagnosis of Schistosoma mansoni infection: parasitological (Kato-Katz), immunological (ELISA) and molecular (real time PCR), and also investigated the sensitivity of each technique in the cure determination after treatment with praziquantel using the water rat Nectomys squamipes, a natural reservoir for S. mansoni, as an experimental model. Two infection laboratory experiments were carried out. The first experiment aimed to observe the evolution of the immunological response in the first moments after infection and in the first months after treatment. The second experiment aimed to compare the efficacy of the three diagnostic techniques after infection and after treatment over a more extended time period. In the first experiment, 44% of the infected animals showed IgG reactivity after two weeks of infection, and 94% were positive based on serology 30 days after infection. The serological IgG titers increased just after infection but decreased gradually after treatment. In the second experiment, 89% of the animals showed positive IgG titers 22 days after infection. Only 68% of the animals showed positive results on the coproscopic diagnostic analysis and 79% did so by qPCR, 50 days after infection. Treated animals showed negative results on coproscopy one month after treatment but remained positive by serology even 12 months after treatment, although showing a decline in immunologic reaction after treatment. By qPCR analysis, all animals showed negative results three months after treatment, except for one animal. The parasitosis can be detected by coproscopy only six weeks after infection, and by serology 14 days after infection. The qPCR was a better diagnostic method for confirming the infection cure of S. mansoni. In early infection, this method was less efficient than serology but was slightly more efficient than the Kato-Katz method. We suggest that the methods should be used in low endemic areas as follows: serology should be used in the initial diagnosis in a population with potential positive cases; subsequently, coproscopy should be used in IgG positive cases to confirm the current infection; and qPCR should be used to evaluate the infection cure after treatment and is also a very valuable tool when there are cases showing positive IgG and negative coproscopy.

Evaluation of polymerase chain reaction as an additional tool for the diagnosis of low-intensity Schistosoma mansoni infection.

The aim of the present study was to evaluate polymerase chain reaction (PCR) as an alternative tool for diagnosing schistosomiasis in individuals with low-level parasite burden from areas of low endemicity or under occasional risk of infection by Schistosoma mansoni. A total of 102 samples were tested in this study using 2 PCR assays utilizing distinct primer pairs. One of the primer pairs was targeted to a highly repeated 121-base pair sequence of S. mansoni, and the other was targeted to Schistosoma 28S rDNA. The samples were divided into 4 groups according to parasite burden of the individual as follows: 16 individuals with schistosomiasis excreting less than 10 eggs per gram of feces (EPG), 18 individuals excreting higher than 10 EPG, 22 individuals with reactive IgG-ELISA against S. mansoni soluble membrane antigen and negative coproscopy, and 46 controls samples including 25 individuals with other intestinal parasites and 21 individuals with negative parasitologic examination. The results obtained with stool samples from individuals with schistosomiasis showed a high sensitivity for PCR as S. mansoni DNA was detected in 91% (31/34) of the samples analyzed. No amplification was observed in 3 stool samples from individuals excreting below 10 EPG. The specificity of the test for both pairs of primers was 100%. In the group of seropositive individuals, S. mansoni DNA was detected in 59% (13/22) of fecal samples, corroborating the serologic results. Overall, PCR can be an important tool for detecting S. mansoni infection in individuals excreting few eggs in feces. Moreover, the determination of the infection through the detection of S. mansoni DNA in stool samples from seropositive individuals represents a new means of confirming the results of IgG-ELISA for schistosomiasis. Therefore, studies in this direction should be encouraged and extended.

[Socio-cultural and ethical factors involved in the diagnosis of schistosomiasis mansoni in an area of low endemicity]. / Fatores sócio-culturais e éticos relacionados com os processos de diagnóstico da esquistossomíase mansônica em área de baixa endemicidade.

Five annual parasitological surveys and one serological survey, respectively based on the Kato-Katz and free sedimentation methods and the Western blot technique, were conducted in Sumidouro, Rio de Janeiro, Brazil, an endemic county for schistosomiasis. Possible influences of the use of these methodologies on social, cultural, and ethical aspects of the study population were also evaluated. Having the opportunity to choose the different techniques was a conclusive issue influencing participation by the population. Prevalence rates of positive results for stool tests were: 11.6% (1995); 8.8% (1996); 12.2% (1998); 5.9% (1999); and 3.2% (2000). In the period during which the serological survey was performed, the use of laboratory testing in association with analysis of clinical data and available data on transmission and treatment generated a diagnostic procedure termed "coproseroepidemiology". This methodology contributed to significant improvements in the accuracy of measurement of local schistosomiasis prevalence, indicating that epidemiological surveillance could help prevent the recurrence of high prevalence rates. The fact that Biomphalaria glabrata was replaced by Melanoides tuberculata in the main transmission focus contributed to a significant decrease in infection rates.

Fatores sócio-culturais e éticos relacionados com os processos de diagnóstico da esquistossomíase mansônica em área de baixa endemicidade / Socio-cultural and ethical factors involved in the diagnosis of schistosomiasis mansoni in an area of low endemicity

Foram realizados em Sumidouro, Rio de Janeiro, Brasil, município endêmico para a esquistossomíase, cinco inquéritos parasitológicos anuais e um sorológico, baseados nas técnicas de Kato-Katz, sedimentação espontânea e western blot. Avaliou-se ainda a interveniência de fatores sociais, culturais e éticos na aplicação de tais métodos junto à comunidade. A possibilidade de opção foi decisiva para a participação das pessoas. As prevalências pela coproscopia foram: 11,6 por cento (1995); 8,8 por cento (1996); 12,2 por cento (1998); 5,9 por cento (1999); 3,2 por cento (2000). No ano de realização da sorologia, os resultados dos exames laboratoriais associados às informações sobre a transmissão, o acompanhamento clínico dos infectados e seus históricos de tratamento, geraram um procedimento diagnóstico nomeado coprosoroepidemiologia que refletiu mais precisamente a prevalência local. Confirmou-se que a vigilância epidemiológica evitaria o recrudescimento da prevalência. A substituição de Biomphalaria glabrata por Melanoides tuberculata no principal foco de transmissão, contribuiu para a queda acentuada dos índices de infecção.

Abundância e infecçäo do molusco Biomphalaria glabrata pelo Schistosoma mansoni no Estado do Rio de Janeiro, Brasil / Abundance and Schistosoma mansoni infection of the snail Biomphalaria glabrata, Brazil

OBJETIVO:Investigar a distribuiçäo espacial, a abundância e os índices de infecçäo natural de Biomphalaria glabrata, hospedeiro intermediário do Schistosoma mansoni, em localidade do Estado do Rio de Janeiro, RJ, Brasil. MÉTODOS:Na localidade de Pamparräo, Município de Sumidouro, RJ, as coletas de moluscos foram realizadas bimestralmente no período de junho de 1991 a novembro de 1995. Foram estabelecidos 23 pontos de coleta ao longo do córrego Pamparräo e três de seus afluentes. Os moluscos capturados foram levados ao laboratório para diagnóstico da infecçäo. Para a análise dos dados, foram usados o coeficiente de Spearman (nível de 0,5 por cento de significância) e o teste de qui-quadrado. RESULTADOS:A abundância populacional de B. glabrata foi bastante variável ao longo do tempo e entre os ambientes amostrados. A maioria dos pontos de coleta apresentou correlaçäo negativa com a pluviosidade. O afluente B destacou-se dos demais corpos d'água por apresentar taxas de infecçäo de B. glabrata elevadas (acima dos 25 por cento em alguns pontos de coleta) e persistentes. Foram encontrados mais moluscos infectados na estaçäo seca do que na chuvosa (?2=20,08; p=0,001). CONCLUSOES:A populaçäo de moluscos foi influenciada negativamente pelo regime de chuvas, principalmente no córrego Pamparräo. A época de estiagem também parece ter favorecido a ocorrência de infecçäo, provavelmente devido ao menor volume de água dos córregos, o que aumentaria as chances de encontro do parasita com seu hospedeiro intermediário