ELENICE MESSIAS DO NASCIMENTO GONCALVES
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Instituto Central, Hospital das Clínicas, Faculdade de Medicina
LIM/03 - Laboratório de Medicina Laboratorial, Hospital das Clínicas, Faculdade de Medicina
LIM/03 - Laboratório de Medicina Laboratorial, Hospital das Clínicas, Faculdade de Medicina
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- Culture isolation and molecular identification of Blastocystis sp. in Brazilian human isolates: preliminary results(2020) MELO, Gessica Baptista de; ROLDAN, William; MALTA, Fernanda de Mello; LESCANO, Susana Angelica Zevallos; CASTILHO, Vera Lucia; GONCALVES, Elenice Messias Do Nascimento; PAULA, Fabiana Martins de; GRYSCHEK, Ronaldo Cesar BorgesBlastocystis sp. is a protist commonly found in stool samples of humans and animals. Biological and genetic factors of this organism remain controversial. The present study aimed to develop and implement the Blastocystis in vitro culture of Brazilian human isolates for routine use. The fecal isolates (n = 20) were maintained in our laboratory by several passages in Pavlova's medium. Cultures were monitored every 72 h by light microscopy. Genomic DNA was extracted to identify the subtypes (STs). In most isolates, the vacuolar form was prevalent. The amoeboid, granular and cystic forms were observed during in vitro cultivation. STs 1, 2, 3. 4 and 7 were identified. Our preliminary results show the generation time and forms present in the in vitro culture of Blastocystis subtypes isolated from Brazilian human isolates. Therefore, we emphasize the use of in vitro culture as a tool in future studies for the better understanding of the biological aspects of Blastocystis sp.
- DIAGNOSIS OF Strongyloides stercoralis INFECTION IN IMMUNOCOMPROMISED PATIENTS BY SEROLOGICAL AND MOLECULAR METHODS(2016) PAULA, Fabiana Martins de; MALTA, Fernanda Mello; CORRAL, Marcelo Andreetta; MARQUES, Priscilla Duarte; GOTTARDI, Maiara; MEISEL, Dirce Mary Correia Lima; YAMASHIRO, Juliana; PINHO, Joao Renato Rebello; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; GRYSCHEK, Ronaldo Cesar Borges; CHIEFFI, Pedro PauloStrongyloidiasis is a potentially serious infection in immunocompromised patients. Thus, the availability of sensitive and specific diagnostic methods is desirable, especially in the context of immunosuppressed patients in whom the diagnosis and treatment of strongyloidiasis is of utmost importance. In this study, serological and molecular tools were used to diagnose Strongyloides stercoralis infections in immunosuppressed patients. Serum and stool samples were obtained from 52 patients. Stool samples were first analyzed by Lutz, Rugai, and Agar plate culture methods, and then by a quantitative real time polymerase chain reaction (qPCR). Serum samples were evaluated by an enzyme-linked immunosorbent assay (ELISA) using a soluble (AS) or a membrane fractions antigen (AM) obtained from alkaline solutions of the filariform larvae of Strongyloides venezuelensis. Of the 52 immunosuppressed patients, three (5.8%) were positive for S. stercoralis by parasitological methods, compared to two patients (3.8%) and one patient (1.9%) who were detected by ELISA using the AS and the AM antigens, respectively. S. stercoralis DNA was amplified in seven (13.5%) stool samples by qPCR. These results suggest the utility of qPCR as an alternative diagnostic tool for the diagnosis of S. stercoralis infection in immunocompromised patients, considering the possible severity of this helminthiasis in this group of patients.
- Characterization of subtypes of Blastocystis sp. isolated from patients with urticaria, Sao Paulo, Brazil(2019) MELO, Gessica Baptista de; MALTA, Fernanda de Mello; MARUTA, Celina Wakisaka; CRIADO, Paulo Ricardo; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; ESPIRITO-SANTO, Maria Cristina de Carvalho do; PAULA, Fabiana Martins de; GRYSCHEK, Ronaldo Cesar BorgesBlastocystis sp. is described as an enteric protist prevalent in fecal samples from humans and animals; its pathogenicity and epidemiology are still controversial. Currently, it has been associated with intestinal diseases such as irritable bowel syndrome and clinical manifestations of allergic skin, such as chronic urticaria. In the context of urticaria, it is still uncertain whether this organism is directly related to the allergic manifestation or just a common component of the intestinal microbiota. This study aimed to evaluate the occurrence and molecular diversity of Blastocystis sp. in individuals with urticaria from a dermatology outpatient clinic, Sao Paulo, Brazil. Fecal samples of 58 patients with urticaria were examined using parasitological methods; and subsequently tested by polymerase chain reaction using Blastocystis-specific primers. The subtypes (STs) and alleles (a) were determined using BLASTn and MLST tools. ST1, ST2, ST3, ST4, ST6 and mixed infection (ST1 + ST3) were identified in the patients with urticaria; ST1 (a4), ST3 (a34 and a36) and ST4 (a42) were the most prevalent. Our molecular analyses allowed an initial description of Blastocystis subtypes in patients with urticaria from Sao Paulo city, Brazil. (C) 2019 Published by Elsevier Ltd on behalf of World Federation of Parasitologists.
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- IMMUNODIAGNOSIS OF HUMAN STRONGYLOIDIASIS: USE OF SIX DIFFERENT ANTIGENIC FRACTIONS FROM Strongyloides venezuelensis PARASITIC FEMALES(2015) CORRAL, Marcelo Andreetta; PAULA, Fabiana Martins de; GOTTARDI, Maiara; MEISEL, Dirce Mary Correia Lima; CASTILHO, Vera Lucia Pagliusi; GONCALVES, Elenice Messias do Nascimento; CHIEFFI, Pedro Paulo; GRYSCHEK, Ronaldo Cesar BorgesThe aim of this study was to evaluate six different antigenic fractions from Strongyloides venezuelensis parasitic females for the immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from S. venezuelensis parasitic females were prepared in phosphate-buffered saline (SSF and SMF, respectively), Tris-HCl (TSF and TMF, respectively), and an alkaline buffer (ASF and AMF, respectively). Serum samples obtained from patients with strongyloidiasis or, other parasitic diseases, and healthy individuals were analyzed by enzyme-linked immunosorbent assay (ELISA). Soluble fractions SSF, TSF, and ASF showed 85.0%, 75.0%, and 80.0% sensitivity and 93.1%, 93.1%, and 87.5% specificity, respectively. Membrane fractions SMF, TMF, and AMF showed 80.0%, 75.0%, and 85.0% sensitivity, and 95.8%, 90.3%, and 91.7% specificity, respectively. In conclusion, the present results suggest that the fractions obtained from parasitic females, especially the SSF and SMF, could be used as alternative antigen sources in the serodiagnosis of human strongyloidiasis.
- 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.