Banca de QUALIFICAÇÃO: PEDRO RAMON DA SILVA AQUINO

Uma banca de QUALIFICAÇÃO de MESTRADO foi cadastrada pelo programa.
DISCENTE : PEDRO RAMON DA SILVA AQUINO
DATA : 19/12/2016
HORA: 09:00
LOCAL: SALA DE AULA II DO PPGCF
TÍTULO:

Sensibilidade analítica e detecção do Trypanosoma cruzi no sangue de indivíduos chagásicos crônicos do Rio Grande do Norte pela Reação em cadeia de Polimerase


PALAVRAS-CHAVES:

Trypanosoma cruzi; Polymerase Chain Reation; Chagas disease, discrete typing units, genetic diversity, Rio Grande do Norte


PÁGINAS: 24
RESUMO:

Objective: In Trypanosoma cruzi infection, Polymerase Chain Reaction (PCR) has been used with great variation in sensitivity in chronic Chagas' patients by region, parasite concentration and greater number of samples. The objective here was to use the Polymerase Chain Reaction for the detection of T. cruzi to know the analytical sensitivity in control samples and in the whole blood of chronically infected individuals from the semi-arid state of Rio Grande do Norte.


Metodology: Blood samples from 68 chronic chagasic individuals aged 22 to 73 years were collected from rural residents with positive serology and submitted to PCR for the detection of T. cruzi kinetoplast DNA (kDNA) with primers 121 and 122 to amplify the 330 bp fragment. The standardization of the positive controls with the isolates RN26, SM73 and SM76, respectively the TcI, TcII and TcIII of Trypanosoma cruzi were amplified with the kDNA PCR and also characterized by genotyping the 3’ region of the 24Sα rRNA, the mitochondrial cytochrome oxidase subunit 2 genes (COII) and the spliced leader intergenic region (SL-IRac).


Results: The standardization of the positive controls was performed in two steps: one in the extraction of the genomic DNA and the second in the DNA concentration in the PCR. On extraction, the parasite dilutions were detected from the concentration of 5x10-4 p / ml for TcII and TcIII, and from 5x10-2 for TcI. With 24Sα of ribosomal DNA (rRNA), SM73 (TcII) obtained 1x10-1 fg / μl, while RN26 (TcI) and SM76 (TcIII), 1 and 10 fg/μL respectively. In the analysis of the spliced leader intergenic region (SL-IRac) genes the analytical sensitivity for TcI and TcIII was 1x10-2 whereas TcII, 1x10-3. The kDNA was positive in 38.2% (26/68) of the chronic chagasic individuals evaluated with positive blood culture in 7.3% (5/68).

Conclusions: The PCR of kDNA was sensitive for the detection of T.cruzi even at very small concentrations of the parasite DNA with amplification differences between DTUs and low frequency of positivity in chronic chagasic probably by genotypic characteristics.


MEMBROS DA BANCA:
Externo ao Programa - 1375489 - ANA CLAUDIA GALVAO FREIRE GOUVEIA
Presidente - 2087759 - ANDRE DUCATI LUCHESSI
Externo ao Programa - 1752367 - PAULO MARCOS DA MATTA GUEDES
Notícia cadastrada em: 15/12/2016 15:13
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