Banca de QUALIFICAÇÃO: STHEPHANIE NASSIF PINHEIRO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : STHEPHANIE NASSIF PINHEIRO
DATA : 04/06/2018
HORA: 14:00
LOCAL: BioME
TÍTULO:

CHARACTERIZATION OF THE 18S RNA GENE IN PROTOZOARS OF THE APICOMPLEXA ROW: AN APPROACH APPLIED TO THE DESIGN OF MOLECULAR MARKERS

PALAVRAS-CHAVES:

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

PÁGINAS: 79
GRANDE ÁREA: Ciências Biológicas
ÁREA: Biologia Geral
RESUMO:

The Apicomplexa phylum comprises protozoa of various genera causing parasitic diseases worldwide such as malaria, toxoplasmosis or opportunistic intestinal disorders. Nowadays, protozoa of medical importance are generally identified by light microscopy, which makes accurate classification difficult, makes diagnosis and prognosis difficult, particularly in cases where infection is low. In this context, the present work aimed to develop an alternative molecular method that allows the identification of a wide range 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 template for screening and species identification in biodiversity studies. From the structural analysis and the ribosomal nucleic acid sequence, sets of primers that interact in conserved regions and flank variable regions of 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 for 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