Padrão de metilação de DNA para fins forenses : análise de células de sangue, sêmen e saliva; e estudo de sensibilidade e especificidade

Detalhes bibliográficos
Ano de defesa: 2015
Autor(a) principal: Silva, Déborah Soares Bispo Santos lattes
Orientador(a): Alho, Clarice Sampaio lattes
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Pontifícia Universidade Católica do Rio Grande do Sul
Programa de Pós-Graduação: Programa de Pós-Graduação em Biologia Celular e Molecular
Departamento: Faculdade de Biociências
País: Brasil
Palavras-chave em Português:
DNA
Área do conhecimento CNPq:
Link de acesso: http://tede2.pucrs.br/tede2/handle/tede/6336
Resumo: DNA obtained from body fluids recovered from crime scenes can be used to identify the donor of the biological material but it cannot reveal the tissue source or the possible age of the donor. DNA methylation is an epigenetic modification involved in transcriptional regulation. It is known that methylation is important in cell differentiation and genomic loci are differentially methylated between tissues. Because of this, different methylation patterns between tissues and cells can provide the basis of an assay for body fluid identification. We also know that the ability to determine the age of the sample donor based on DNA would be a powerful tool for forensic investigation. Human aging is associated with epigenetic modifications such as DNA methylation. Several studies have investigated biomarkers for aging which can be used to track donor age, presenting practical implications in forensic analysis. Two genes previously found to be DNA methylation age-associated, NPTX2 and GRIA2, were tested for prediction of age in saliva and blood samples. Both age markers were hypermethylated with the increase of age. The epigenetic predicted age was calculated for both markers, with an average difference of 6.9 years between estimated and observed ages for GRIA2 marker, and an average difference of 9.2 years for NPTX2 marker. Other studies investigated biomarkers for identification of bodyfluids. The most common body fluids found at crime scenes are blood, semen and saliva. A set of epigenetic markers, cg-06379435, ZC3H12D and BCAS4, which produce unique and specific patterns of DNA methylation, can be used to identify these body fluid types. However, to ensure the efficiency of these epigenetic markers, developmental validation studies need to be performed to determine the conditions and limitations of this new tool for forensic analysis. When testing the markers for body fluid identification using different organisms, we did obtain positive results for certain primate samples, however all other tested species were negative. The lowest concentration consistently detected varied from 0.1 to 10ng, depending on the locus. The method also proved to be effective when inhibitors were present in the samples or when samples were degraded by heat. In the case of mixtures, the overall methylation values varied in a consistent and predictable manner when multiple cell types were present in the same sample. Overall, the search of age markers candidates can be an important key in tracking suspects/victims in forensic investigations, and the ability to identify the biological sample using DNA would be a powerful tool. Epigenetic markers will provide the forensic community with new and improved methods to interpret the crime scene.