Comparação da taxa de detecção do DNA do EBV em amostras de saliva total e células exfoliadas de pacientes com leucoplasia bucal através da nPCR

• Autor(a) principal: Franco, Carolline Vitor Alves [UNESP]
• Data de Publicação: 2016
• Tipo de documento: Trabalho de conclusão de curso
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/155238; http://www.athena.biblioteca.unesp.br/exlibris/bd/capelo/2017-04-25/000881197.pdf
• topic: Leucoplasia bucal; Herpesvirus humano 4; Carcinoma de células escamosas; Leukoplakia, Oral

The aim of this essay was to compare the detection rate of Epstein-Barr virus (EBV) DNA in total saliva samples with samples of exfoliated cells, obtained through the use of cytobrush of patients with oral leukoplakia, through n-PCR. In the laboratory phase, the DNA was extracted from the samples according to the instructions of the DNA extraction Kit QIAamp® DNA mini kit and all samples were submitted to spectrophotometry to verify presence and integrity of the DNA. Subsequently the PCRs were performed for β-globin, with primers PCO3 and PCO4 (260pb). To verify the presence of human DNA, an analysis was performed using 2% agarose gel electrophoresis. n-PCR was performed with initiator oligonucleotides for the EBV's DNA. All samples were tested in duplicate and the n-PCR products were subjected to 8% polyacrylamide gel electrophoresis. It was verified the existence of association between variables of both groups studied by applying statistical tests (Chi-square or Fisher's exact Test) with a significance level of 5%. Saliva and exfoliated cells samples collected in the laboratory phase for DNA extraction and PCR for β-globin, which was positive for all samples, showing a good quality DNA. EBV's reaction standardization revealed that 80% of the samples (24 patients) of cytobrush were positive for the presence of the virus. In this analysis the prevalence was in males who presented positivity in 84% of cases (16 patients). As for the analysis of the saliva samples, the positivity was 77% (23 patients) also maintaining the prevalence of male with 69.5% of cases (16 patients). However the research showed no statistical significance in the comparison of the detection rate of Epstein-Barr virus (EBV) DNA in samples evaluated of patients with oral leukoplakia