Navegando por Palavras-chave "Genetic Diversity"
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- ItemSomente MetadadadosAnálise da Diversidade Genética em Agentes da Esporotricose Usando Marcadores Aflp (Amplified Fragment Length Polymorphism)(Universidade Federal de São Paulo (UNIFESP), 2020-03-10) Carvalho, Jamile Ambrosio De [UNIFESP]; Camargo, Zoilo Pires De [UNIFESP]; Universidade Federal de São PauloSporothrix is a thermodimorphic fungus found in the environment which causes sporotrichosis, a subcutaneous mycosis that can be transmitted to humans directly from contaminated soil or plants through traumatic inoculum from fungal propagules or through bites and scratches from contaminated cats. The genus is divided into two clades, a clinical clade represented by S. brasiliensis, S. schenckii, S. globosa, and S. luriei, and an environmental clade represented by species rarely described as human pathogens. Considering the ongoing epidemic occurring in Brazil, mostly caused by S. brasiliensis, it is necessary to use techniques to explore diversity within the species. The existing techniques for this type of study have high costs, we aimed to standardize the AFLP technique for application in Sporothrix. AFLP is based on genomic DNA digestion using one or more restriction enzymes and selective amplification of the fragments generated using selective molecular primers with the addition of one or more nucleotides to the 3' region of these primers. The technique has been used for several microorganisms and has advantages as not requiring the knowledge of the sequence of isolates. To reduce costs and time, firstly, an in silico AFLP was performed to predict the fragments that would be generated and thus to choose the nucleotide combinations that presented more efficiency to separate the isolates by species. For this, we use two softwares: AFLPinSilico and ISIF. The two softwares were compatible with each other and both revealed six combinations that showed good results for studies in Sporothrix. These combinations were tested in vitro and all obtained good results, but only three were selected to continue the study with 188 isolates of Sporothrix from different geographic regions. In general, the study presented three new markers that demonstrated efficiency in separating Sporothrix by species and accessing the intraspecific diversity in the species of the clinical clade, clarifying questions of the expansion of S. brasiliensis.
- ItemRestritoLevantamento das espécies de Bostrychia dos manguezais de Bertioga, Santos - Cubatão e Complexo Estuarino-Lagunar de Iguape-Cananéia, com base em DNA barcoding(Universidade Federal de São Paulo, 2022-02-07) Santos, Juliana Moreira dos [UNIFESP]; Milstein, Daniela [UNIFESP]; http://lattes.cnpq.br/7442898844707725; http://lattes.cnpq.br/9069947770491880; Universidade Federal de São Paulo (UNIFESP)As macroalgas associadas aos mangues são organismos de suma importância ecológica nos ecossistemas estuarinos. Nos manguezais do Brasil, destacam-se as espécies do gênero Bostrychia. Estudos sobre a sua diversidade, quando baseados apenas em abordagens morfológicas, podem ser imprecisos pois os caracteres empregados para a identificação de espécies nem sempre são informativos. Além disso, algumas espécies são crípticas, o que muitas vezes leva a uma identificação duvidosa. No entanto, quando dados moleculares são associados a caracteres morfológicos, a delimitação em nível de espécie fica mais acurada. O presente projeto utilizou a técnica de DNA barcoding para identificar as espécies do gênero Bostrychia que ocorrem nos manguezais de Bertioga, Santos-Cubatão e Cananéia-Iguape. Os resultados obtidos com o sequenciamento dos marcadores moleculares da mitocôndria (COI-5P) e cloroplasto (rbcL), indicaram a ocorrência de quatro espécies distintas (B. montagnei, B. pilulifera, B. radicans e Bostrychia sp). Sendo, B. pilulifera uma nova ocorrência para os manguezais do Estado de São Paulo. A espécie identificada como Bostrychia sp. não obteve
- ItemSomente MetadadadosMolecular identification of Histoplasma capsulatum using rolling circle amplification(Wiley-Blackwell, 2016) Furuie, Jason L.; Sun, Jiufeng; do Nascimento, Mariana M. F.; Gomes, Renata R.; Waculicz-Andrade, Caroline E.; Sessegolo, Gisele C.; Rodrigues, Anderson M. [UNIFESP]; Galvao-Dias, Maria A.; de Camargo, Zoilo P. [UNIFESP]; Queiroz-Telles, Flavio; Najafzadeh, Mohammad J.; de Hoog, Sybren G.; Vicente, Vania A.Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatumDNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.
- ItemSomente MetadadadosMolecular identification of Histoplasma capsulatum using rolling circle amplification(Wiley-Blackwell, 2016) Furuie, Jason L.; Sun, Jiufeng; do Nascimento, Mariana M. F.; Gomes, Renata Rodrigues; Waculicz-Andrade, Caroline E.; Sessegolo, Gisele C.; Rodrigues, Anderson Messias [UNIFESP]; Galvao-Dias, Maria Adelaide; Camargo, Zoilo Pires de [UNIFESP]; Queiroz-Telles, Flavio; Najafzadeh, Mohammad J.; de Hoog, Sybren G.; Vicente, Vania Aparecida; Universidade Federal de São Paulo (UNIFESP)Histoplasmosis is a systemic fungal disease that occurs worldwide, causing symptomatic infection mostly in immunocompromised hosts. Etiological agent is the dimorphic fungus, Histoplasma capsulatum, which occurs in soil contaminated with bird or bat droppings. Major limitation in recognition of H. capsulatum infections is the low awareness, since other diseases may have similar symptomatology. The molecular methods have gained importance because of unambiguous diagnostic ability and efficiency. The aim of this study was to develop and evaluate a padlock probe in view of rolling circle amplification (RCA) detection method which targets ITS (Internal Transcribed Spacer) rDNA of H. capsulatum enabling rapid and specific detection of the fungus in clinical samples. Two padlock probes were designed and one of these (HcPL2) allowed specific amplification of H. capsulatumDNA while no cross-reactivity was observed with fungi used as negative controls. This method proved to be effective for H. capsulatum specific identification and demonstrated to be faster than the traditional method of microbiological identification.