Banca de QUALIFICAÇÃO: STHEPHANIE NASSIF PINHEIRO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : STHEPHANIE NASSIF PINHEIRO
DATA : 23/02/2018
HORA: 15:00
LOCAL: DEPTO DE BIOQUÍMICA - CENTRO DE BIOCIÊNCIAS
TÍTULO:

CARATERIZAÇÃO DO GENE 18S rRNA EM PROTOZOÁRIOS DO TAXA APICOMPLEXA: UMA ABORDAGEM APLICADA AO DESENHO DE MARCADORES MOLECULARES

PALAVRAS-CHAVES:

18S rRNA. semi nested-PCR. Apicomplexan. PCR.

PÁGINAS: 50
GRANDE ÁREA: Ciências Biológicas
ÁREA: Biologia Geral
RESUMO:

The Apicomplexa taxa comprises protozoa of various genera causing parasitic diseases worldwide such as malaria, toxoplasmosis or opportunistic intestinal disorders. Even today, protozoa of medical importance are generally identified by light microscopy, which makes accurate classification difficult and makes diagnosis and prognosis difficult, particularly in cases where the infection is low. In this context, the present work aimed to develop an alternative molecular method that allows the identification of a wide variety of protozoa of the Apicomplexa taxa. Thus, a primer system was developed for use in a semi-nested PCR (Polymerase Chain Reaction) reaction. The investigated target for primer design was the 18S rDNA region, as it is a widely used target for screening and species identification in biodiversity studies. From the structural analysis and the ribosomal nucleic acid sequence, sets of primers flanking variable regions in the gene were designed. The efficiency of each set of primers was evaluated by in silico PCR and the generated amplicons were evaluated. A set of primers was selected which, when used in a nested fashion, can generate ~ 166 amplicons with distinct sequences, which can be used to discriminate genera and species of the Apicomplexa taxa by difference in the size of amplicons generated in agarose gel and species by sequencing (Sanger method or Next Gen Sequencing). The proposed method was validated in vitro and its efficiency in the identification of some protozoan species of medical interest was confirmed. After further validation steps this method can be used for initial screening in cases of suspected parasitosis and also for parasite species determination.

MEMBROS DA BANCA:
Presidente - 1880243 - DANIEL CARLOS FERREIRA LANZA
Interno - 1513597 - JOAO PAULO MATOS SANTOS LIMA