DNA extraction and molecular analysis of non-tumoral liver, spleen, and brain from autopsy samples: the effect of formalin fixation and paraffin embedding

DNA extraction and molecular analysis of non-tumoral liver, spleen, and brain from autopsy samples: the effect of formalin fixation and paraffin embedding

Author Funabashi, Karina Silva Autor UNIFESP Google Scholar
Barcelos, Denise Autor UNIFESP Google Scholar
Visona, Iria Autor UNIFESP Google Scholar
Silva, Marcelo Souza e Autor UNIFESP Google Scholar
Sousa, Maria Luiza Almeida Prado Oliveira e Autor UNIFESP Google Scholar
Franco, Marcello Fabiano de Autor UNIFESP Google Scholar
Miazato Iwamura, Edna Sadayo Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Inst Criminalist São Paulo
Abstract The use of molecular biology in combination with morphological analysis is increasing because of the treatments by target therapies. However, to improve the methods for obtaining DNA for molecular analyses from formalin-fixed, paraffin-embedded (FFPE) tissue is a challenge. the aim of this study was to evaluate the DNA extracted from FFPE tissue blocks (non-tumoral liver, spleen, and brain), obtained from autopsy, 8-24h post mortem, using three methods of DNA extraction. PCR of the beta-actin (136pb) and human amelogenin (AMEL 212-218 bp/106-112 bp) genes, as well as short tandem repeat (STR) (100-400 bp fragments), reported in forensic scientific analysis, was performed to evaluate the effectiveness of the methods of DNA extraction. We used 28 archived (1 and 5 years) and 12 recent autopsy cases. the commercial kit showed reproducible and consistent results in the PCR amplification of the beta-actin and AMEL genes and in analysis by SIR used in forensic analysis. This is the first report using non-tumoral samples from FFPE autopsy tissues, comparing the three most common methods of DNA extraction and using the SIR previously described in forensics. Our study has clarified the challenges for pathologists in applying the molecular biology approach in combination with methods suited for morphology, which must be improved. the data provided here should be used in other molecular studies in FFPE samples. (C) 2012 Elsevier GmbH. All rights reserved.
Keywords Molecular pathology
Autopsy
Formalin-fixed
Paraffin-embedded
PCR
STR
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Grant number FAPESP: 2008/11233-8
Date 2012-01-01
Published in Pathology Research and Practice. Jena: Elsevier Gmbh, Urban & Fischer Verlag, v. 208, n. 10, p. 584-591, 2012.
ISSN 0344-0338 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 584-591
Origin http://dx.doi.org/10.1016/j.prp.2012.07.001
Access rights Closed access
Type Article
Web of Science ID WOS:000311072700003
URI http://repositorio.unifesp.br/handle/11600/34462

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