MARIA CRISTINA CARVALHO DO ESPIRITO SANTO
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Unidades Organizacionais
LIM/06 - Laboratório de Imunopatologia da Esquistossomose e outras Parasitoses, Hospital das Clínicas, Faculdade de Medicina
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conferenceObject DETECTION OF SCHISTOSOMA MANSONI SPOROCYST STAGE IN BIOMPHALARIA GLABRATA MOLLUSK IN EXPERIMENTAL CONDITIONS(2018) CASOTTI, Marcia; TUAN, Roseli; GOMES, Michele; LUNA, Expedito Albuquerque; DIAS-NETO, Emmanuel; PAULA, Fabiana; PINHO, Joao Rebello; CARRILHO, Flair; GRYSCHEK, Ronaldo Borges; ESPIRITO-SANTO, MariaconferenceObject MOLECULAR CHARACTERIZATION OF THE LARVAL PHASE OF SCHISTOSOMA MANSONI IN BIOMPHALARIA GLABRATA MOLLUSKS UNDER EXPERIMENTAL CONDITIONS(2017) CASOTTI, Marcia Oliveira; TUAN, Roseli Tuan; GOMES, Michele; DIAS-NETO, Emmanuel; PINHO, Joao Renato Rebello; PAULA, Fabiana Martins; CARRILHO, Flair Jose Carrilho Jose; LUNA, Expedito Jose Albuquerque; GRYSCHEK, Ronaldo Cesar Borges Borges; ESPIRITO-SANTO, Maria Cristina- TWO SEQUENTIAL PCR AMPLIFICATIONS FOR DETECTION OF Schistosoma mansoni IN STOOL SAMPLES WITH LOW PARASITE LOAD(2012) ESPIRITO-SANTO, Maria Cristina Carvalho do; ALVARADO-MORA, Monica Viviana; PINTO, Pedro Luiz Silva; CARRILHO, Flair Jose; PINHO, Joao Renato Rebello; GRYSCHEK, Ronaldo Cesar BorgesSchistosomiasis constitutes a major public health problem, with an estimated 200 million individuals infected worldwide and 700 million people living in risk areas. In Brazil there are areas of high, medium and low endemicity. Studies have shown that in endemic areas with a low prevalence of Schistosoma infection the sensitivity of parasitological methods is clearly reduced. Consequently diagnosis is often impeded due to the presence of false-negative results. The aim of this study is to present the PCR reamplification (Re-PCR) protocol for the detection of Schistosoma mansoni in samples with low parasite load (with less than 100 eggs per gram (epg) of feces). Three methods were used for the lysis of the envelopes of the S. mansoni eggs and two techniques of DNA extraction were carried out. Extracted DNA was quantified, and the results suggested that the extraction technique, which mixed glass beads with a guanidine isothiocyanate/phenol/chloroform (GT) solution, produced good results. PCR reamplification was conducted and detection sensitivity was found to be five eggs per 500 mg of artificially marked feces. The results achieved using these methods suggest that they are potentially viable for the detection of Schistosoma infection with low parasite load.
conferenceObject REVALENCE OF SCHISTOSOMA MANSONI INFECTION AND OTHER PARASITIC DISEASES IN PERIPHERAL AREAS OF BARRA MANSA, RIO DE JANEIRO, BRAZIL(2017) ESPIRITO-SANTO, Maria Cristina C.; CHIEFFI, Pedro Paulo; PAULA, Fabiana Martins de; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; ORBAN, Magali; PINHO, Joao Renato Rebello; LUNA, Expedito Jose de Albuquerque; GRYSCHEK, Ronaldo Cesar Borges- Molecular detection of prepatent Schistosoma mansoni infection in Biomphalaria glabrata snail vectors(2020) CASOTTI, Marcia Oliveira; GRYSCHEK, Ronaldo Cesar Borges; PAULA, Fabiana Martins de; GOMES-GOUVEA, Michele; PINHO, Joao Renato Rebello; TUAN, Roseli; DIAS-NETO, Emmanuel; LUNA, Expedito Jose de Albuquerque; ESPIRITO-SANTO, Maria Cristina Carvalho doApproximately 240 million people worldwide are infected by Schistosoma. In Brazil, one of the main intermediate hosts of this parasite is Biomphalaria glabrata snails. The early detection of larval stages in intermediate hosts is an important challenge to public health. but it also represents an opportunity as a new alternative to indicate earlier natural infections before cercariae differentiation and emergence. In this context, we demonstrated that PCR amplification of a 28S gene fragment from the parasite does demonstrate S. mansoni infection in snails 14 days post infection. This conventional polymerase chain reaction amplified clear bands and was able to detect parasitic infection in the intermediate host B. glabrata under experimental conditions. However, we reinforce that this approach requires deeper investigations and further comparisons to confirm its specificity and sensitivity in earlier time points after miracidia infection. This approach has relevant potential as an effective molecular-based strategy for the monitoring of schistosomiasis transmission.
- Comparative Study of the Accuracy of Different Techniques for the Laboratory Diagnosis of Schistosomiasis Mansoni in Areas of Low Endemicity in Barra Mansa City, Rio de Janeiro State, Brazil(2015) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; PINTO, Pedro Luiz Silva; SANCHEZ, Maria Carmen Arroyo; DIAS-NETO, Emmanuel; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; CHIEFFI, Pedro Paulo; LUNA, Expedito Jose de Albuquerque; PINHO, Joao Renato Rebello; CARRILHO, Flair Jose; GRYSCHEK, Ronaldo Cesar BorgesSchistosomiasis constitutes a major public health problem, with an estimated 200 million people infected worldwide. Many areas of Brazil show low endemicity of schistosomiasis, and the current standard parasitological techniques are not sufficiently sensitive to detect the low-level helminth infections common in areas of low endemicity (ALEs). This study compared the Kato-Katz (KK); Hoffman, Pons, and Janer (HH); enzyme-linked immunosorbent assay-(ELISA-) IgG and ELISA-IgM; indirect immunofluorescence technique (IFT-IgM); and qPCR techniques for schistosomiasis detection in serum and fecal samples, using the circumoval precipitin test (COPT) as reference. An epidemiological survey was conducted in a randomized sample of residents from five neighborhoods of Barra Mansa, RJ, with 610 fecal and 612 serum samples. ELISA-IgM(21.4%) showed the highest positivity and HH and KK techniques were the least sensitive (0.8%). All techniques except qPCR-serum showed high accuracy (82-95.5%), differed significantly from COPT in positivity (P < 0.05), and showed poor agreement with COPT. Medium agreement was seen with ELISA-IgG (Kappa = 0.377) and IFA (Kappa = 0.347). Parasitological techniques showed much lower positivity rates than those by other techniques. We suggest the possibility of using a combination of laboratory tools for the diagnosis of schistosomiasis in ALEs.
- Evaluation of real-time PCR assay to detect Schistosoma mansoni infections in a low endemic setting(2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; DIAS-NETO, Emmanuel; BOTELHO-LIMA, Livia Souza; MOREIRA, Joao Paulo; AMORIM, Maria; PINTO, Pedro Luiz Silva; HEATH, Ashley R.; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; LUNA, Expedito Jose de Albuquerque; CARRILHO, Flair Jose; PINHO, Joao Renato Rebello; GRYSCHEK, Ronaldo Cesar BorgesBackground: Schistosomiasis constitutes a major public health problem, and 200 million people are estimated to be infected with schistosomiasis worldwide. In Brazil, schistosomiasis has been reported in 19 states, showing areas of high and medium endemicity and a wide range of areas of low endemicity (ALE). Barra Mansa in Rio de Janeiro state has an estimated prevalence of 1%. ALE represent a new challenge for the helminth control because about 75% of infected individuals are asymptomatic and infections occur with a low parasite load (<100 eggs per gram of feces), causing a decrease in sensitivity of stool parasitological techniques, which are a reference for the laboratory diagnosis of this helminth. The objective of this study was to evaluate the performance of a TaqMan quantitative polymerase chain reaction (qPCR) technique in serum and feces DNA samples using the techniques of Kato-Katz (KK), Hoffman, Pons and Janer (HH) as references, during an epidemiological survey using fecal samples and sera from randomized residents from an ALE. Methods: A cross-sectional study conducted from April to December 2011 using a probabilistic sampling that collected 572 fecal and serum samples. The laboratory diagnostic techniques used were: KK, HH and qPCR ( feces and serum). Results: We obtained the following results using the different diagnostic techniques: KK and HH, 0.9% (n = 5); qPCR-feces, 9.6% (n = 55); and qPCR-serum, 1.4% (n = 8). The qPCR-feces presented the highest positivity, whereas the techniques of HH and KK were the least sensitive to detect infections (0.8%). Compared to HH and KK, qPCR-feces showed a statistically significant difference in positivity (p < 0.05), although with poor agreement. Conclusion: The positivity rate presented by the qPCR approach was far higher than that obtained by parasitological techniques. The lack of adequate surveillance in ALE of schistosomiasis indicates a high possibility of these areas being actually of medium and high endemicity. This study presents a control perspective, pointing to the possibility of using combined laboratory tools in the diagnosis of schistosomiasis in ALE.
- Detection of Schistosoma mansoni infection by TaqMan (R) Real-Time PCR in a hamster model(2014) ESPIRITO-SANTO, Maria Cristina Carvalho; ALVARADO-MORA, Monica Viviana; PINTO, Pedro Luiz Silva; BRITO, Thales de; BOTELHO-LIMA, Livia; HEATH, Ashley Richard; AMORIM, Maria Galli; DIAS-NETO, Emmanuel; CHIEFFI, Pedro Paulo; PINHO, Joao Renato Rebello; CARRILHO, Flair Jose; LUNA, Expedito Jose Albuquerque; GRYSCHEK, Ronaldo Cesar BorgesAn experimental study in hamsters was performed to evaluate the capability for detecting Schistosoma mansoni DNA in serum and fecal samples during the pre and post-egg-laying periods of infection using TaqMan (R) Real-Time PCR system (qPCR), was compared with the circumoval precipitin test (COPT) and the Kato-Katz technique, especially among individuals with low parasitic burden. Twenty-four hamsters were infected with cercariae. Three hamsters were sacrificed per week under anesthesia, from 7 days post infection (DPI) up to 56 DPI. A serum sample and a pool of feces were collected from each hamster. The presence of S. mansoni eggs in fecal samples was evaluated by Kato-Katz method and in the hamsters gut-by histopathology. Detection of S. mansoni DNA was performed using qPCR and S. mansoni antibody using COPT. The first detection of eggs in feces by Kato-Katz method and S. mansoni DNA in feces by qPCR occurred 49 DPI. Nevertheless, S. mansoni DNA was detected in serum samples from 14 up to 56 DPI. COPT was positive at 35 DPI. The results not only confirm the reliability of S. mansoni DNA detection by qPCR, but also demonstrate that serum is a trustworthy source of DNA in the pre patent infection period.
- Detection of Schistosoma mansoni Antibodies in a Low-Endemicity Area Using Indirect Immunofluorescence and Circumoval Precipitin Test(2014) ESPIRITO-SANTO, Maria Cristina Carvalho do; PINTO, Pedro Luiz; GARGIONI, Cybele; ALVARADO-MORA, Monica Viviana; CASTILHO, Vera Lucia Pagliusi; PINHO, Joao Ranato Rebello; LUNA, Expedito Jose de Albuquerque; GRYSCHEK, Ronaldo Cesar BorgesParasitological diagnostic methods for schistosomiasis lack sensitivity, especially in regions of low endemicity. The objective of this study was to determine the prevalence of Schistosoma mansoni infections by antibody detection using the indirect immunofluorescence assay (IFA-IgM) and circumoval precipitin test (COPT). Serum samples of 572 individuals were randomly selected. The IFA-IgM and COPT were used to detect anti-S. mansoni antibodies. Of the patients studied, 15.9% (N = 91) were IFA-IgM positive and 5.1% (N = 29) had COPT reactions (P < 0.001 by McNemar's test). Immunodiagnostic techniques showed higher infection prevalence than had been previously estimated. This study suggests that combined use of these diagnostic tools could be useful for the diagnosis of schistosomiasis in epidemiological studies in areas of low endemicity.