Banca de DEFESA: PEDRO RAMON DA SILVA AQUINO

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
DISCENTE : PEDRO RAMON DA SILVA AQUINO
DATA : 25/08/2017
HORA: 14:00
LOCAL: SALA 2 DO PPgCF
TÍTULO:

Analytical sensitivity and detection of Trypanosoma cruzi in the blood of chronic Chagas' individuals from Rio Grande do Norte by Polymerase Chain Reaction

PALAVRAS-CHAVES:

Chagas' disease, parasitological diagnosis; Polimerase Chain Reaction; Trypanosoma cruzi, DTUs.

PÁGINAS: 86
GRANDE ÁREA: Ciências da Saúde
ÁREA: Farmácia
RESUMO:

Trypanosoma cruzi is subdivided into six Discrete Typing Units (DTUs), TcI-TcVI. In the semiarid of Rio Grande do Norte (RN), TcI, TcII and TcIII were identified in triatomines and humans. This study aimed to evaluate the analytical sensitivity of the Polymerase Chain Reaction (PCR) for the detection of Trypanosoma cruzi in TcI, TcII and TcIII and in the whole blood of chronically infected individuals from the semiarid region of Rio Grande do Norte. Dilutions of parasites and DNA from DTUs I, II and III isolated from individuals with positive blood culture and 100 blood samples from residents of rural communities of different municipalities of the RN with reactive serology were submitted to PCR for the detection of kDNA and the microsatellite region (SatDNA) of T. cruzi. The analytical sensitivity of the DTU genotyping was performed using the markers: subunit II mitocondrial gene citocromo oxidase (CO II) , divergent domain of the ribosomal DNA 24S gene (rRNA) and intergenic spacer of T. cruzi (SL-IR). Analytical sensitivity assays demonstrated amplification variations between DTUs with better performance of kDNA in relation to SatDNA. In samples from chronic chagasic individuals, 36% amplified the 330 bp fragment of kDNA, while 19% of these same samples amplified the 188 bp fragment of SatDNA. All samples positive for SatDNA were also in kDNA. In the PCR positive samples, 75.0% were from untreated individuals and 25.0% from treated individuals. The analysis using CO II, rRNA and SL-IR showed amplification variations between the DTUs, but were not effective in the dilutions of 0.5 to 0.00005 parasite/mL and also in the genotyping of blood samples from chronic chagasic individuals. The lower positivity found in the state is probably related to the genetic constitution of the parasite, as well as factors such as circulating genotype, etiological treatment, virulence or susceptibility to immune response that influence parasitemia and, consequently, detection by PCR.

MEMBROS DA BANCA:
Presidente - 1218940 - ANTONIA CLAUDIA JACOME DA CAMARA
Externo ao Programa - 1752367 - PAULO MARCOS DA MATTA GUEDES
Externo à Instituição - HELENA KEIKO TOMA - UFRJ