PRISCILLA DUARTE MARQUES FONSECA
Projetos de Pesquisa
Unidades Organizacionais
LIM/06 - Laboratório de Imunopatologia da Esquistossomose e outras Parasitoses, Hospital das Clínicas, Faculdade de Medicina
10 resultados
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- Immunoreactivity of proteins within 30-40 kDa range during the acute and the recovery phases in rats experimentally infected with Strongyloides venezuelensis(2020) FONSECA, Priscilla Duarte Marques; CORRAL, Marcelo Andreeta; MEISEL, Dirce Mary C. Lima; LEVI, Debora; NASCIMENTO, Rafael Correa; CASTRO-BORGES, William; GRYSCHEK, Ronaldo; COSTA-CRUZ, Julia Maria; PAULA, Fabiana Martins deIn experimental infection with Strongyloides venezuelensis, the acute and recovery phases can be distinguished, unlike human infections caused by Strongyloides stercoralis. The objective of this study was to evaluate the production of anti-Strongyloides IgG antibodies and the recognition of immunogenic protein bands during the acute and the recovery phases in rats experimentally infected with S. venezuelensis. Rats were infected subcutaneously with 400 or 4,000 S. venezuelensis infective larvae. The acute phase was characterized by elimination of a large number of eggs in the faeces on days 6-14 post infection; the recovery phase was characterized by the resolution of the infection between days 30 and 35 post infection. Differences in IgG levels were observed in the acute and the recovery phases. Different antigenic fractions were recognized in both phases of infection. It is concluded that proteins within the 30-40 kDa range are immunoreactive markers for both the acute and the recovery phases in rats experimentally infected with S. venezuelensis, particularly using membrane antigen.
- 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.
- 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.
- Shotgun proteomics of Strongyloides venezuelensis infective third stage larvae: Insights into host-parasite interaction and novel targets for diagnostics(2020) FONSECA, Priscilla D. M.; CORRAL, Marcelo A.; COSENZA-CONTRERAS, Miguel; MEISEL, Dirce M. C. L.; MELO, Gessica B.; ANTUNES, Milena M. S.; SANTO, Maria C. E.; GRYSCHEK, Ronaldo C. B.; COSTA-CRUZ, Julia M.; CASTRO-BORGES, William; PAULA, Fabiana M.Strongyloides venezuelensis is an important alternative source of antigen for the serologic diagnosis of human strongyloidiasis. Proteomics techniques applied to the analysis of the protein content of infective third stage larvae (iL3) of S. venezuelensis provide a powerful tool for the discovery of new candidates for immunodiagnosis. This study presents an overview of the protein iL3 S. venezuelensis focusing on the diagnosis of strongyloidiasis. A total of 877 proteins were identified by shotgun proteomics. Many of these proteins are involved in different cellular processes, metabolic as well as structural maintenance. Our results point to a catalog of possible diagnostic targets for human strongyloidiasis and highlight the need for evaluation of uncharacterized proteins, especially the proteins within the CAP domain, transthyretin, and BTPI inhibitor domains, as a repertoire as yet unexplored in the context of strongyloidiasis diagnostic markers. We believe that the protein profile presented in this shotgun analysis extends our understanding of the protein composition within the Strongyloides genus, opening up new perspectives for research on biomarkers that may help with the diagnosis of human strongyloidiasis. Data are available via ProteomeXchange with identifier PXD013703.
- Shotgun proteomics of Strongyloides venezuelensis infective third stage larvae: Insights into host-parasite interaction and novel targets for diagnostic (vol 235, 111249, 2020)(2020) FONSECA, Priscilla D. M.; CORRAL, Marcelo A.; COSENZA-CONTRERAS, Miguel; MEISEL, Dirce M. C. L.; MELO, Gessica B.; ANTUNES, Milena M. S.; SANTO, Maria C. E.; GRYSCHEK, Ronaldo C. B.; COSTA-CRUZ, Julia M.; CASTRO-BORGES, William; PAULA, Fabiana M.
- 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.
conferenceObject DIFFERENTIAL EXPRESSION OF MEMBRANE AND MEMBRANE-BOUND PROTEINS FROM FILARIFORM LARVAE AND ADULT FEMALE OF STRONGYLOIDES VENEZUELENSIS(2017) PAULA, Fabiana M.; CORRAL, Marcelo A.; MARQUES, Priscilla D.; MEISEL, Dirce Mary C.; COSTA-CRUZ, Julia Maria; SANTO, Maria Cristina Espirito; CAMPOS, Jonatan M.; MATTEI, Bruno; BORGES, William Castro; GRYSCHEK, Ronaldo Cesar- Evaluation of targets for Strongyloides genus specific molecular diagnosis in experimental strongyloidiasis(2021) NASCIMENTO, Rafael C.; MELO, Gessica B.; FONSECA, Priscilla D. M.; GRYSCHEK, Ronaldo C. B.; PAULA, F. M.Strongyloides venezuelensis has been used in different experimental studies, such as those aimed at the evaluation of diagnostic techniques for human strongyloidiasis, mainly the molecular diagnosis. In this study, three regions (genus, 18S and 28S targets) of Strongyloides ribosomal DNA were evaluated for the molecular diagnosis of experimental strongyloidiasis. Rats were infected subcutaneously with 400 or 4000 S. venezuelensis infective larvae (400iL3 and 4000iL3), and kept for 35 days. Fecal samples were collected daily to count eggs per gram of feces (EPG) and to perform the polymerase chain reaction (PCR). Egg count started on the 5th day post-infection (pi) and ended on days 33 and 34 pi, in 400iL3 and 4000iL3 groups, respectively. Based in EPG, fecal samples were selected from days 2, 5, 8, 11, 15, 23 and 35 pi for DNA extraction; PCR (genus, 18S and 28S); and sequencing. The PCR-28S products showed higher values of identity (95-100%) in the database with the Strongyloides sequences. Therefore, it is possible to reinforce the application of PCR-28S in the diagnosis of experimental and human strongyloidiasis.
- 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.
- Diagnosis of the strongyloid nematode Strongyloides venezuelensis in experimentally infected rats(2016) MARQUES, P. D.; MALTA, F. M.; MEISEL, D. M. C. L.; CORRAL, M. A.; PINHO, J. R.; COSTA-CRUZ, J. M.; CHIEFFI, P. P.; GRYSCHEK, R. C. B.; PAULA, F. M.Strongyloides venezuelensis is an intestinal nematode of rats, frequently used as a model for studying human and animal strongyloidiasis. In the present study, we evaluated parasitological, serological and molecular methods for the diagnosis of experimental S. venezuelensis in rats, Rattus norvegicus. Blood and faecal samples were collected and analysed up to 60 days post infection (pi) with adult worm recovery occurring from 5 to 45 days pi. Using an enzyme-linked immunosorbent assay (ELISA), serum levels of IgG antibodies increased up to 28 days pi, thereafter decreasing by day 60 pi. Polymerase chain reaction (PCR) assays detected S. venezuelensis DNA in faecal samples of rats from 5 to 21 days pi. The present study therefore represents the first step towards improving the diagnosis of experimental strongyloidiasis.