Banca de QUALIFICAÇÃO: MARIA FERNANDA BEZERRA DE SOUZA

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
STUDENT : MARIA FERNANDA BEZERRA DE SOUZA
DATE: 28/05/2021
TIME: 14:00
LOCAL: Videoconferência - Link para acesso: https://meet.google.com/yox-idcu-eif
TITLE:

MOLECULAR MARKERS DEVELOPED FOR IDENTIFICATION OF Plasmodium sp.

KEY WORDS:

Apicomplexan- Plasmodium-PCR-18S-MSP1

PAGES: 85
BIG AREA: Ciências Biológicas
AREA: Bioquímica
SUMMARY:

Early, the Plasmodium genus had only 4 species related to human malaria: P. vivax, P. falciparum, P. malariae and P. ovale. With the advancement of molecular tools and bioinformatics, there has been an increase in the number of species that cause this disease in humans such as P. knowlessi, P. cynolmolgi and P. simiun. The polymerase chain reaction (PCR) has been considered the most effective among the detection techniques. Most of the PCR protocols already described for the identification of Plasmodium sp. targets the ribosomal RNA 18S subunit gene.However, primers for the identification of P. vivax, P. falciparum, P. malariae and P. ovale generally cross-react with other species, species that can also cause malaria in humans. The pattern of evolution of the 18S gene in the apicomplex group is marked by copies that resemble each other by species, by genus (Plasmodium) and no apicomplex group. Despite the great efficiency of Primers for 18S rDNA, this target does not enable differentiation of Plasmodium species with great efficiency.Thus, a complementary system was developed, based on another molecular marker, for the identification of Plasmodium species. Available sequences were selected for six species that cause malaria in humans (Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae, Plasmodium ovale, Plasmodium knowlessi and Plasmodium cynomolgi). Thus, the sequences of the several of genes were investigated. The validation of the molecular marker for Plasmodium was based on the specificity and the number of sequences available for all species. Among them, the gene chosen for the design of the primers was MSP1 (merozoite surface protein). The Plasmodium identification system was built of way to produce amplicons of different size for each species, making it possible to identify the species without the need for sequencing the amplicon. Then, due to the particularity of the 18S, a nested-PCR already proposed for the detection of apicomplex was validated for an initial screening. And a nested-PCR system was developed for P. vivax and P. falciparum that were tested in vitro.It was possible to verify that the systems for the two most recurrent species in Brazil, show sensitivity and specificity. By verifying the amplification of both samples extracted from cell culture and the DNA of patients. Specificity within the Plasmodium genus has also been confirmed. Through the verification of non-amplification from samples of Plasmodium berghei and it was also observed that there is no cross amplification between samples of P. vivax and P. falciparum.

BANKING MEMBERS:
Interno - 1046922 - LEONARDO CAPISTRANO FERREIRA
Externa à Instituição - PATRICIA DE ABREU MOREIRA - UFOP
Presidente - 2213126 - VALTER FERREIRA DE ANDRADE NETO