LUMENA PEREIRA MACHADO SIQUEIRA
Projetos de Pesquisa
Unidades Organizacionais
Instituto Central, Hospital das Clínicas, Faculdade de Medicina
LIM/53 - Laboratório de Micologia, Hospital das Clínicas, Faculdade de Medicina
LIM/53 - Laboratório de Micologia, Hospital das Clínicas, Faculdade de Medicina
7 resultados
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- Comparing the phenotypic, genotypic, and proteomic identification of Trichosporon species: A globally emerging yeast of medical importance(2021) LARA, Bruna Rossini; CAMARGO, Bruno Braidotti de; PAULA, Claudete Rodrigues; LEITE JUNIOR, Diniz Pereira; GARCES, Hans Garcia; ARNONI, Mariana Volpe; SILVEIRA, Monica; GIMENES, Viviane Mazo Favero; SIQUEIRA, Lumena Pereira Machado; TAKAHASHI, Juliana Possatto Fernandes; MELHEM, Marcia de Souza Carvalho; RICHINI-PEREIRA, Virginia Bodelao; ANVERSA, Lais; RUIZ, Luciana da SilvaTrichosporon spp. are widely distributed in the nature, comprising species that inhabit different ecological niches and can be found in the water, soil, and body surface of animals and humans. Such microorganisms have been classically associated with superficial infections; however, in the last decades, they have also been related to disseminated infections in immunocompromised patients, behaving as opportunistic agents, which demands rapid and accurate species identification for efficient therapy. Concordance level between the traditional phenotypic method and the molecular technique (gold standard) in the identification of all 59 Trichosporon samples was 59.3%. Identification concordance between MALDI-TOF spectrometry and the molecular technique was 71.2%. No isolate of environmental origin was identifiable by MALDI-TOF mass spectrometry (MS), and 100% of such environmental isolates were discordant for IGS region sequencing and phenotypic characterization. Both comparisons evidenced greatest concordance in the identification of T. asahii. The species T. debeurmannianum, T. dermatis, T. venhuisii and T. insectorum were not properly identified by both MALDI-TOF MS and the phenotypic technique. MALDI-TOF MS, in particular, seems to be appropriate to investigate yeasts of the genus Trichosporon; however, database updates are still necessary, especially for species that are not common in the clinical routine. With the aim of helping understand the aspects involved in early and accurate diagnosis of infections caused by this opportunistic agent, the present study compared the phenotypic, molecular (IGS region) and mass-spectrometry (MALDI-TOF) identification of 59 yeasts of the genus Trichosporon which had clinical and environmental origin and were kept in a mycology collection.
- First report of tinea corporis caused by Arthroderma benhamiae in Brazil(2019) FREITAS, Roseli Santos de; FREITAS, Thais Helena Proenca de; SIQUEIRA, Lumena Pereira Machado; GIMENES, Viviane Mazo Favero; RENARD, GilArthroderma benhamiae is a zoophilic dermathophyte that can cause highly inflammatory tinea corporis and tinea capitis in humans. This is the first report of a patient with dermatophytosis caused by A. benhamiae in Brazil. The lesion was an erythematous, annular plaque on the lumbar region that appeared few weeks after playing with a street cat in a 19-month-old girl. Initial presumed diagnosis was tinea corporis caused by Microsporum canis. Topical treatments were ineffective and the patient required systemic treatment with griseofulvin. Mycological diagnosis was inconclusive: morphological differentiation between M. canis and Trichophyton benhamiae may be difficult, especially when the latter present yellow colonies. The etiological agent was identified only by ITS sequencing of the isolates aligned with reference strains to A. benhamiae. This report highlights the importance of ITS sequencing in the identification of isolates from some cases of dermatophytosis, because conventional morphological diagnosis may result in misdiagnosis of the agent and delay proper treatment.
- A neglected disease. Human sporotrichosis in a densely populated urban area in Sao Paulo, Brazil: clinical-epidemiological and therapeutic aspects(2022) BITTENCOURT, Amanda Azevedo; OYAFUSO, Luiza Keiko Matsuka; CAVALIN, Roberta Figueiredo; PALHARES, Renata Bacic; BENARD, Gil; GIMENES, Viviane Mazo Favero; NEGRO, Gilda Maria Barbaro Del; SIQUEIRA, Lumena Pereira Machado; XAVIER, Roseli Santos de Freitas; LOPES-BEZERRA, Leila M.; BUCCHERI, Renata; LINDOSO, Jose Angelo LaulettaHuman sporotrichosis is caused by different Sporothrix species; however, Sporothrix brasiliensis is the main species, usually related to cat transmission in urban areas. A retrospective descriptive study was conducted at the Institute of Infectology Emilio Ribas from 2010 to 2018. Demography, clinical, diagnostic, and therapeutic data were obtained from medical records. Polymerase chain reaction of the calmodulin gene was performed to identify Sporothrix species. In addition, to evaluate the spread of the disease across Sao Paulo metropolitan region, TerraView version 4.2.2 software was used for geocoding cases according to residence addresses. Kernell's maps using QGIS software version 2.16.3 were constructed to determine the concentration of cases. Results: 260 cases of sporotrichosis were diagnosed between 2010 and 2018. We observed a 700% increment in the number of human cases in the 2016-2018 triennium compared with the 2013-2015 triennium. Female adults with a median age of 46 years old were the predominant infected group associated with cats' exposition at home care, although the age range of all patients was 01 to 86 years old. The main epidemiological risk of acquiring sporotrichosis was contact with cats, reported by 96.5% of the patients. Molecular identification showed that most of the tested isolates were Sporothrix brasiliensis. Lymphocutaneous form was observed in 59.2% and fixed cutaneous form in 37.5% of the patients. Regarding treatment, itraconazole was the main drug used (94.2%) with a cure rate of 98.8%. We observed an important spread of human sporotrichosis involving cat transmission caused by Sporothrix brasiliensis in a densely populated area of Sao Paulo state. These results are important to alert clinicians and dermatologists about the occurrence and progression of a neglected tropical disease in an urban area and the urgent necessity to include sporotrichosis as a differential diagnosis in the clinical investigation routine.
- Polysaccharide diversity in VNI isolates of Cryptococcus neoformans from Roraima, Northern Brazil(2019) PUPO, Halan D. Dal; SENA, Bianca A. G.; REIS, Flavia C. G.; MACHADO, Lumena; FORTES, Silvana T.; ALMEIDA JUNIOR, Joao N. de; GODINHO, Rodrigo M. C.; RODRIGUES, Marcio L.Species of the Cryptococcus genus comprise environmental, encapsulated fungal pathogens that cause lethal meningitis in immunosuppressed individuals. In humans, fungal uptake of hypocapsular Cryptococcus by macrophages was associated with high fungal burden in the cerebrospinal fluid and long-term patient survival. On the basis of the key role of the cryptococcal capsule in disease, we analyzed the diversity of capsular structures in 23 isolates from pigeon excreta collected in the cities of Boa Vista, Bonfim and Pacaraima, in the state of Roraima (Northern Brazil). All isolates were identified as Cryptococcus neoformans (VNI genotype) by MALDI-TOF mass spectrometry. Through a combination of fluorescence microscopy, flow cytometry, ELISA and spectrophotometric methods, each isolate was characterized at the phenotypical level, which included measurements of growth rates at 30 and 37 degrees C, pigmentation, cell body size, capsular dimensions, serological reactivity, urease production and ability to produce extracellular glucuronoxylomannan (GXM), the main capsular component of C. neoformans. With the exception of melanization, a formidable diversity was observed considering all parameters tested in our study. Of note, hyper and hypo producers of GXM were identified, in addition to isolates with hyper and hypo profiles of reactivity with a polysaccharide-binding monoclonal antibody. Capsular dimensions were also highly variable in the collection of isolates. Extracellular GXM production correlated positively with capsular dimensions, urease activity and cell size. Unexpectedly, GXM concentrations did not correlate with serological reactivity with the cryptococcal capsule. These results reveal a high diversity in the ability of environmental C. neoforrnans to produce capsular components, which might impact the outcome of human cryptococcosis.
- Evaluating VITEK MS for the identification of clinically relevant Aspergillus species(2020) AMERICO, Fernanda M.; SIQUEIRA, Lumena P. Machado; NEGRO, Gilda Maria B. Del; GIMENES, Viviane M. Favero; TRINDADE, Mario Roberto S.; MOTTA, Adriana L.; FREITAS, Roseli Santos de; ROSSI, Flavia; COLOMBO, Arnaldo L.; BENARD, Gil; ALMEIDA JUNIOR, Joao N. deAspergillus spp. identification has become more relevant in clinical practice since azole-resistant cryptic species have been related to invasive fungal infections. Conventional morphologic identification is not able to discriminate Aspergillus species, and DNA sequencing is not feasible for clinical laboratories. MALDI-TOF mass spectrometry is an emergent technology that has been explored to provide fast and accurate identification of microorganisms, including clinically relevant moulds. However, only a few studies have explored the platform VITEK MS for the identification of Aspergillus species. Hence, we provided additional data regarding the performance of the VITEK MS system for the identification of Aspergillus species, including azole-resistant ones. We also improved the RUO system by adding additional spectral profiles from well-identified Aspergillus strains belonging to different noncryptic and cryptic species. The IVD library correctly identified 91.6% of the organisms at genus and section level, and 84.7% at species level, including the azole-resistant Aspergillus lentulus and Aspergillus calidoustus. The organisms belonging to Aspergillus cryptic species had only 31.2% of correct species identification. The RUO library plus our in-house SuperSpectra correctly identified 100% of the organisms at genus and section level and 91.6% at species level. Among organisms belonging to Aspergillus cryptic species, 68.7% had correct species identification. Some closely related Aspergillus cryptic species showed similar spectral profiles and were difficult to be differentiated.
- Evaluation of Vitek MS for Differentiation of Cryptococcus neoformans and Cryptococcus gattii Genotypes(2019) SIQUEIRA, Lumena P. Machado; GIMENES, Viviane M. Favero; FREITAS, Roseli Santos de; MELHEM, Marcia de Souza Carvalho; BONFIETTI, Lucas Xavier; JR, Afonso Rafael da Silva; SANTOS, Leticia B. Souza; MOTTA, Adriana L.; ROSSI, Flavia; BENARD, Gil; JR, Joao N. de AlmeidaCryptococcus neoformans and Cryptococcus gattii are the main pathogenic species of invasive cryptococcosis among the Cryptococcus species. Taxonomic studies have shown that these two taxa have different genotypes or molecular types with biological and ecoepidemiological peculiarities. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been proposed as an alternative method for labor-intensive methods for C. neoformans and C. gattii genotype differentiation. However, Vitek MS, one of the commercial MALDI-TOF MS instruments, has not been yet been evaluated for this purpose. Thus, we constructed an in-house database with reference strains belonging to the different C. neoformans (VNI, VNII, VNIII, and VNIV) and C. gattii (VGI, VGII, VGIII, and VGIV) major molecular types by using the software Saramis Premium (bioMerieux, Marcy-l'Etoile, France). Then, this new database was evaluated for discrimination of the different genotypes. Our in-house database provided correct identification for all C. neoformans and C. gattii genotypes; however, due to the intergenotypic mass spectral similarities, a careful postanalytic evaluation is necessary to provide correct genotype identification.
- Significance of Aspergillus spp. isolation in defining cases of COVID-19 Associated Pulmonary Aspergillosis - CAPA(2023) COCIO, Tiago Alexandre; SIQUEIRA, Lumena Pereira Machado; RICILUCA, Katie Cristina Takeuti; GIMENES, Viviane Mazo Favero; ANDRADE, Tania Sueli de; BENARD, Gil; MARTINEZ, Roberto; BOLLELA, Valdes RobertoCOVID-19-Associated Pulmonary Aspergillosis (CAPA) is a relatively common complica-tion in patients with severe forms of the disease caused by the SARS-CoV-2 virus. Diag-nosing and confirming CAPA is challenging. In this study, Aspergillus spp. isolation in respiratory specimens from patients with COVID-19 was evaluated for identifying cases of CAPA. In 2020-2021, 17 Aspergillus spp. were isolated from 15 COVID-19 patients admit-ted to a university hospital in Brazil. Patient records were retrospectively reviewed to obtain clinical-epidemiological data and other markers of Aspergillus spp. infection and then compared with the ECMM/ISHAM criteria for defining CAPA. Probable CAPA was defined in 5/10 patients, who had Aspergillus spp. isolated from Bronchoalveolar Lavage (BAL) or a positive galactomannan blood test. Additionally, anti-Aspergillus antibodies were detected in two of these patients, during active or follow-up phases of CAPA. In another seven patients with Aspergillus spp. isolated from tracheobronchial aspirate or sputum, CAPA was presumed, mainly due to deterioration of clinical conditions and new lung imaging suggestive of fungal infection. Antifungal agents to control CAPA, particu-larly voriconazole, were used in 9/15 cases. In cases of probable CAPA and remaining patients, clinical conditions and comorbidities were similar, with lethality being high, at 60% and 71%, respectively. The number of CAPA cases defined by scientific criteria was lower than that assumed in the clinical context. This was largely due to the lack of BAL collection for fungal culture and the non-intensive use of other markers of invasive asper-gillosis. The isolation of Aspergillus spp. in different respiratory specimens should alert clinicians to the diagnosis of CAPA.