FABIANA MARTINS DE PAULA
Projetos de Pesquisa
Unidades Organizacionais
LIM/06 - Laboratório de Imunopatologia da Esquistossomose e outras Parasitoses, Hospital das Clínicas, Faculdade de Medicina
14 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 14
- Molecular diagnosis of strongyloidiasis in tropical areas: a comparison of conventional and real-time polymerase chain reaction with parasitological methods(2015) PAULA, Fabiana Martins de; MALTA, Fernanda de Mello; MARQUES, Priscilla Duarte; SITTA, Renata Barnabe; PINHO, Joao Renato Rebello; GRYSCHEK, Ronaldo Cesar Borges; CHIEFFI, Pedro PauloThis study aimed to evaluate the use of conventional polymerase chain reaction (cPCR) and real-time quantitative PCR (qPCR) in the diagnosis of human strongyloidiasis from stool samples in tropical areas. Stool samples were collected from individuals and were determined to be positive for Strongyloides stercoralis (group I), negative for S. stercoralis (group II) and positive for other enteroparasite species (group III). DNA specific to S. stercoralis was found in 76.7% of group I samples by cPCR and in 90% of group I samples by qPCR. The results show that molecular methods can be used as alternative tools for detecting S. stercoralis in human stool samples in tropical areas.
- 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 PHYLOGENETIC ANALYSIS OF BLASTOCYSTIS SPP. ISOLATES IN CLINICAL STOOL SAMPLES FROM BRAZIL(2017) GRYSCHEK, Ronaldo; MELO, Gessica; PAULA, Fabiana; MALTA, Fernanda; MARUTA, Celina; CRIADO, Paulo; MAGRI, Marcello; CASTILHO, Vera; GONCALVES, Elenice- Molecular diagnosis of Strongyloides stercoralis among transplant candidates(2018) PAULA, Fabiana M.; MALTA, Fernanda M.; MARQUES, Priscilla D.; MELO, Gessica B.; CORRAL, Marcelo A.; GOTTARDI, Maiara; PINHO, Joao R. R.; GONCALVES, Elenice M. N.; CASTILHO, Vera L. P.; PIERROTTI, Ligia C.; ABDALA, Edson; COSTA, Silvia F.; CHIEFFI, Pedro P.; GRYSCHEK, Ronaldo C. B.Strongyloidiasis can occur without any symptoms or as a potentially fatal hyperinfection or disseminated infection, principally in immunosuppressed patients. Our study aimed to evaluate the application of conventional polymerase chain reaction (cPCR) and real-time PCR (qPCR). Polymerase chain reaction (PCR) and real-time PCR (qPCR) targeting the 18S rRNA gene for detection of Strongyloides stercoralis infection among transplant candidates were applied in stool samples obtained from 150 transplant candidates, preliminarily analyzed by parasitological methods. S.stercoralis larvae were visualized in 15/150 (10.0%) transplant candidates by parasitological methods. DNA from S.stercoralis was amplified in 26/150 (17.3%) and 49/150 (32.7%) stool samples of transplant candidates, using cPCR and qPCR, respectively. The results suggest that molecular methods, especially qPCR, should be used as an additional tool for diagnostic of S.stercoralis infection among transplant candidates.
- PARASITOLOGICAL AND MOLECULAR DIAGNOSIS IN EXPERIMENTAL Strongyloides venezuelensis INFECTION(2013) PAULA, Fabiana Martins; SITTA, Renata Barnabe; MALTA, Fernanda Mello; GOTTARDI, Maiara; CORRAL, Marcelo Andreetta; GRYSCHEK, Ronaldo Cesar Borges; CHIEFFI, Pedro PauloStrongyloides venezuelensis is a parasitic nematode of rats which is frequently used as a model to study human and animal strongyloidiasis. The aim of this study was to evaluate the correlation between parasitological and molecular diagnosis in Strongyloides venezuelensis infection. PCR assays were used to detect S. venezuelensis DNA in fecal samples obtained from experimentally infected Rattus norvegicus. The results showed a higher sensitivity of the PCR assay in detecting the infection compared to parasitological methods.
- Toxocara DNA amplification in serum and tissue samples in BALB/c mice(2021) FONSECA, Gabriela Rodrigues e; MELO, Gessica Baptista de; PAULA, Fabiana Martins de; MALTA, Fernanda Mello; GRYSCHEK, Ronaldo Cesar Borges; LESCANO, Susana Angelica ZevallosToxocariasis is still a neglected parasitic disease worldwide and much about its biology and diagnosis has yet to be understood. The migration of third stage larvae via bloodstream suggests a potential use of molecular tools in diagnosis as well to deepen the knowledge about its migration behaviors. Conventional PCR was applied in serum and tissue samples from BALB/c mice infected with 5 and 500 embryonated eggs. Blood samples were collected at 15, 30, 60, 90 and 120 days post-infection. Organs were excised at 170 days post infection. There was no DNA amplification in serum samples in any group or day post-infection; contrarily, tissue samples showed DNA amplification. These results also support a continuous larval migration after and/or simultaneously with the neurotropic-myotropic phase. Thus, molecular tools might be useful as a differential diagnosis method, but do not replace immunodiagnostics techniques.
conferenceObject GENETIC DIVERSITY OF BLASTOCYSTIS SUBTYPES IN PATIENTS WITH CHRONIC URTICARIA(2017) PAULA, Fabiana M.; MELO, Gessica B.; MALTA, Fernanda M.; MARUTA, Celina W.; CRIADO, Paulo R.; CASTILHO, Vera Lucia P.; GONCALVES, Elenice M. N.; SANTO, Maria Cristina Espirito; GRYSCHEK, Ronaldo Cesar- Molecular and Immnune Diagnosis: Further Testing for Human Strongyloidiasis(2018) BOSQUI, Larissa R.; MARQUES, Priscilla D.; MELO, Gessica B. de; GONCALVES-PIRES, Maria do Rosario F.; MALTA, Fernanda M.; PAVANELLI, Wander R.; CONCHON-COSTA, Ivete; COSTA-CRUZ, Julia M.; PAULA, Fabiana M.; COSTA, Idessania N.Detection of Strongyloides stercoralis larvae is particularly challenging because only a small number of larvae are released into the feces, regardless of infection stage. Our objective was to apply conventional polymerase chain reaction (PCR) to the detection of S. stercoralis DNA in feces samples to evaluate its performance in samples of patients with strongyloidiasis and compare results with those of immunodiagnosis. Stool, serum, and saliva samples were collected from each individual (n = 48) at the clinic hospital of the State University of Londrina, Brazil, for parasitological, immunological, and molecular tests. Stool samples were processed via parasitological methods. Serum samples were used for immunoglobulin G (IgG) detection and saliva samples for IgA detection by ELISA. For amplification by conventional PCR, two different primers were used: species specific (101 bp) and genus specific (392 bp). The results showed that 34 (97.1%) of the 35 copro-positive individuals for S. stercoralis were positive for serum IgG and 19 (54.3%) were positive for salivary IgA. Regarding molecular analysis, both primers (species and genus specific) demonstrated positivity in 100% of the samples, which was confirmed by sequencing the positive samples. Complementary examinations of the parasitological method demonstrated excellent results in the context of the diagnosis of strongyloidiasis, especially in asymptomatic patients with irregular larval release in the feces.