GILDA MARIA BARBARO DEL NEGRO
Projetos de Pesquisa
Unidades Organizacionais
LIM/53 - Laboratório de Micologia, Hospital das Clínicas, Faculdade de Medicina
48 resultados
Resultados de Busca
Agora exibindo 1 - 10 de 48
- Candida blankii: an emergent opportunistic yeast with reduced susceptibility to antifungals(2018) ALMEIDA JR., Joao Nobrega de; CAMPOS, Silvia V.; THOMAZ, Danilo Y.; THOMAZ, Luciana; ALMEIDA, Renato K. G. de; NEGRO, Gilda M. B. del; GIMENES, Viviane F.; GRENFELL, Rafaella C.; MOTTA, Adriana L.; ROSSI, Flavia; BENARD, Gil
- Identification and antifungal susceptibility of Candida species isolated from the urine of patients in a university hospital in Brazil(2017) LIMA, Glaucia Moreira Espindola; NUNES, Maina de Oliveira; CHANG, Marilene Rodrigues; TSUJISAKI, Rosianne Assis de Sousa; NUNES, Joslaine de Oliveira; TAIRA, Cleison Ledesma; THOMAZ, Danilo Yamamoto; NEGRO, Gilda Maria Barbaro Del; MENDES, Rinaldo Poncio; PANIAGO, Anamaria Mello MirandaThe aim of this study was to identify Candida spp. isolated from candiduria episodes at a tertiary hospital in the Midwest region of Brazil, and to determine their susceptibility profiles to antifungal compounds. From May 2011 to April 2012, Candida spp. isolated from 106 adult patients with candiduria admitted to the University Hospital of the Federal University of Mato Grosso do Sul were evaluated. Both, species identification and susceptibility testing with fluconazole-FLC, voriconazole-VRC, and amphotericin B-AmB were carried out using the Vitek 2. To discriminate species of the C. parapsilosis complex, a RAPD-PCR technique using the RPO2 primer was performed. From the total of 106 isolates, 42 (39.6%) C. albicans and 64 (60.4%) Candida non-albicans (CNA) -33 C. tropicalis, 18 C. glabrata, 5 C. krusei, 4 C. parapsilosis sensu stricto, 2 C. kefyr, 1 C. lusitaniae, and 1 C. guilliermondii were identified. All isolates were susceptible to AmB and VRC, whereas all C. glabrata isolates presented either resistance (5.6%) or dose-dependent susceptibility (94.4%) to FLC. The study of Candida spp. and their resistance profiles may help in tailoring more efficient therapeutic strategies for candiduria.
- Rapid identification of moulds and arthroconidial yeasts from positive blood cultures by MALDI-TOF mass spectrometry(2016) ALMEIDA JR., Joao N. de; SZTAJNBOK, Jaques; SILVA JUNIOR, Afonso Rafael da; VIEIRA, Vinicius Adriano; GALASTRI, Anne Layze; BISSOLI, Leandro; LITVINOV, Nadia; NEGRO, Gilda Maria Barbaro Del; MOTTA, Adriana Lopes; ROSSI, Flavia; BENARD, GilMoulds and arthroconidial yeasts are potential life-threatening agents of fungemia in immunocompromised patients. Fast and accurate identification (ID) of these pathogens hastens initiation of targeted antifungal therapy, thereby improving the patients' prognosis. We describe a new strategy that enabled the identification of moulds and arthroconidial yeasts directly from positive blood cultures by MALDI-TOFmass spectrometry (MS). Positive blood cultures (BCs) with Gram staining showing hyphae and/or arthroconidia were prospectively selected and submitted to an in-house protein extraction protocol. Mass spectra were obtained by Vitek MS (TM) system, and identifications were carried out with in the research use only (RUO) mode with an extended database (SARAMIS (TM) [v.4.12] plus in-house database). Fusarium solani, Fusarium verticillioides, Exophiala dermatitidis, Saprochaete clavata, and Trichosporon asahii had correct species ID by MALDI-TOF MS analysis of positive BCs. All cases were related to critically ill patients with high mortality fungemia and direct ID from positive BCs was helpful for rapid administration of targeted antifungal therapy.
- First report of a clinical isolate of Candida haemulonii in Brazil(2012) ALMEIDA JR., Joao Nobrega de; MOTTA, Adriana Lopes; ROSSI, Flavia; ABDALA, Edson; PIERROTTI, Ligia Camera; KONO, Adriana Satie Goncalves; DIZ, Maria Del Pilar Estevez; BENARD, Gil; NEGRO, Gilda Maria Barbaro Del
- Serological Diagnosis of Paracoccidioidomycosis: High Rate of Inter-laboratorial Variability among Medical Mycology Reference Centers(2014) VIDAL, Monica Scarpelli Martinelli; NEGRO, Gilda Maria Barbaro Del; VICENTINI, Adriana Pardini; SVIDZINSKI, Teresinha Inez Estivalet; MENDES-GIANNINI, Maria Jose; ALMEIDA, Ana Marisa Fusco; MARTINEZ, Roberto; CAMARGO, Zoilo Pires de; TABORDA, Carlos Pelleschi; BENARD, GilBackground: Serological tests have long been established as rapid, simple and inexpensive tools for the diagnosis and follow-up of PCM. However, different protocols and antigen preparations are used and the few attempts to standardize the routine serological methods have not succeeded. Methodology/Principal findings: We compared the performance of six Brazilian reference centers for serological diagnosis of PCM. Each center provided 30 sera of PCM patients, with positive high, intermediate and low titers, which were defined as the ""reference'' titers. Each center then applied its own antigen preparation and serological routine test, either semiquantitative double immunodifusion or counterimmmunoelectrophoresis, in the 150 sera from the other five centers blindly as regard to the ""reference'' titers. Titers were transformed into scores: 0 (negative), 1 (healing titers), 2 (active disease, low titers) and 3 (active disease, high titers) according to each center's criteria. Major discordances were considered between scores indicating active disease and scores indicating negative or healing titers; such discordance when associated with proper clinical and other laboratorial data, may correspond to different approaches to the patient's treatment. Surprisingly, all centers exhibited a high rate of ""major'' discordances with a mean of 31 (20%) discordant scores. Alternatively, when the scores given by one center to their own sera were compared with the scores given to their sera by the remaining five other centers, a high rate of major discordances was also found, with a mean number of 14.8 sera in 30 presenting a discordance with at least one other center. The data also suggest that centers that used CIE and pool of isolates for antigen preparation performed better. Conclusion: There are inconsistencies among the laboratories that are strong enough to result in conflicting information regarding the patients' treatment. Renewed efforts should be promoted to improve standardization of the serological diagnosis of PCM.
- Accuracy of the QuantiFERON-TB Gold in Tube for diagnosing tuberculosis in a young pediatric population previously vaccinated with Bacille Calmette-Guerin(2014) VALLADA, Marcelo Genofre; OKAY, Thelma Suely; DEL NEGRO, Gilda Maria B.; ANTONIO, Claudio Amaral; YAMAMOTO, Lidia; RAMOS, Sonia Regina T. S.Objective: To evaluate the accuracy of an interferongamma release assay (QuantiFERON-TB Gold in Tube) for diagnosing Mycobacterium tuberculosis infection in a young pediatric population. Methods: 195 children previously vaccinated with BCG were evaluated, being 184 healthy individuals with no clinical or epidemiological evidence of mycobacterial infection, and 11 with Mycobacterium tuberculosis infection, according to clinical, radiological, and laboratory parameters. A blood sample was obtained from each child and processed according to the manufacturer's instructions. The assay performance was evaluated by a Receiver Operating Characteristic (ROC) curve. Results: In the group of 184 non-infected children, 130 (70.6%) were under the age of four years (mean age of 35 months). In this group, 177 children (96.2%) had negative test results, six (3.2%) had indeterminate results, and one (0.5%) had a positive result. In the group of 11 infected children, the mean age was 58.5 months, and two of them (18%) had negative results. The ROC curve had an area under the curve of 0.88 (95%CI 0.82-0.92; p<0.001), disclosing a predictive positive value of 81.8% for the test (95%CI 46.3-97.4). The assay sensitivity was 81.8% (95%CI 48.2-97.2) and the specificity was 98.8% (95%CI 96-99.8). Conclusions: In the present study, the QuantiFERON-TB Gold in Tube performance for diagnosing M. tuberculosis infection was appropriate in a young pediatric population.
- A Real Time PCR strategy for the detection and quantification of Candida albicans in human blood(2020) BUSSER, Felipe Delatorre; COELHO, Vivian Caso; FONSECA, Claudia de Abreu; NEGRO, Gilda Maria Barbaro Del; SHIKANAI-YASUDA, Maria Aparecida; LOPES, Marta Heloisa; MAGRI, Marcello Mihailenko Chaves; FREITAS, Vera Lucia Teixeira deCandidemia is a significant cause of bloodstream infections (BSI) in nosocomial settings. The identification of species can potentially improve the quality of care and decrease human mortality. Quantitative PCR (qPCR) was evaluated for Candida albicans detection using culture suspensions containing C. albicans, spiked human blood. the cloned qPCR target fragment (ITS2 region) and the results of these assays were compared. The assays showed a good detection limit: C. albicans DNA extracted from yeast (sensitivity 0.2 CFU/mu L), spiked human blood (sensitivity 10 CFU/mL), and cloned fragment of ITS2 region (sensitivity 20 target copies/mu L). The efficiency of ITS2 fragment-qPCR ranged from 89.67 to 97.07, and the linearity (R-2) of the standard curve ranged from 0.992 to 0.999. The results showed that this ITS2-qPCR has a great potential as a molecular prototype model for the development of a test to be applied in clinical practice, greatly reducing the time of candidemia diagnosis, which is extremely important in this clinical setting.
- Is the S405F mutation in Candida albicans ERG11 gene sufficient to confer resistance to fluconazole?(2014) OLIVEIRA-CARVALHO, V.; NEGRO, G. M. B. Del
- Trichosporon inkin as an Emergent Pathogen in Patients With Severe Pemphigus(2015) ALMEIDA JUNIOR, Joao Nobrega de; OLIVEIRA, Renata Buccheri de; DUARTE, Amaro; MOTTA, Adriana Lopes; ROSSI, Flavia; FIGUEIREDO, Dulce Sachiko Yamamoto de; NEGRO, Gilda Maria Barbaro Del; AOKI, Valeria; MARUTA, Celina Wakisaka; SANTI, Claudia Giuli; BENARD, GilIMPORTANCE To our knowledge, these are the first reports of bloodstream infections by Trichosporon inkin in patients with pemphigus. OBSERVATIONS Trichosporon inkin, a novel organism causing bloodstream infection, was detected in 2 patients with pemphigus. An elderly man with pemphigus foliaceus died despite treatment with liposomal amphotericin B, 3mg/kg/d, and a young girl with pemphigus vulgaris responded to treatment with voriconazole, 8mg/kg/d, for 24 days. One of the T inkin isolates had a minimal inhibitory concentration of 2mg/L against amphotericin B, suggesting resistance to the drug. CONCLUSIONS AND RELEVANCE Delayed suspicion of invasive infection by T inkin may result in a poor outcome in patients with severe forms of pemphigus. This opportunistic infection is highly refractory to conventional potent antifungal treatment.
- Usefulness of matrix-assisted laser desorption ionisation-time-of-flight mass spectrometry for identifying clinical Trichosporon isolates(2014) ALMEIDA JUNIOR, J. N. de; FIGUEIREDO, D. S. Y.; TOUBAS, D.; NEGRO, G. M. B. Del; MOTTA, A. L.; ROSSI, F.; GUITARD, J.; MORIO, F.; BAILLY, E.; ANGOULVANT, A.; MAZIER, D.; BENARD, G.; HENNEQUIN, C.Trichosporon spp. have recently emerged as significant human pathogens. Identification of these species is important, both for epidemiological purposes and for therapeutic management, but conventional identification based on biochemical traits is hindered by the lack of updates to the species databases provided by the different commercial systems. In this study, 93 strains, or isolates, belonging to 16 Trichosporon species were subjected to both molecular identification using IGS1 gene sequencing and matrix-assisted laser desorption ionisation-time-of-flight (MALDI-TOF) analysis. Our results confirmed the limits of biochemical systems for identifying Trichosporon species, because only 27 (36%) of the isolates were correctly identified using them. Different protein extraction procedures were evaluated, revealing that incubation for 30 min with 70% formic acid yields the spectra with the highest scores. Among the six different reference spectra databases that were tested, a specific one composed of 18 reference strains plus seven clinical isolates allowed the correct identification of 67 of the 68 clinical isolates (98.5%). Although until recently it has been less widely applied to the basidiomycetous fungi, MALDI-TOF appears to be a valuable tool for identifying clinical Trichosporon isolates at the species level.