Advanced search
Start date
Betweenand
Related content
(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

Full text
Author(s):
Funabashi, Karina Silva [1] ; Barcelos, Denise [1] ; Visona, Iria [1] ; Silva, Marcelo Souza e [1] ; Sousa, Maria Luiza Almeida Prado Oliveira e [1, 2] ; de Franco, Marcello Fabiano [1] ; Miazato Iwamura, Edna Sadayo [1]
Total Authors: 7
Affiliation:
[1] Univ Fed Sao Paulo, Escola Paulista Med, Dept Pathol, EPM UNIFESP, BR-04023062 Sao Paulo - Brazil
[2] Inst Criminalist Sao Paulo, Lab Biol & Bioquim, Sao Paulo - Brazil
Total Affiliations: 2
Document type: Journal article
Source: PATHOLOGY RESEARCH AND PRACTICE; v. 208, n. 10, p. 584-591, 2012.
Web of Science Citations: 15
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. (AU)

FAPESP's process: 08/11233-8 - DNA extraction and analysis of amelogenin in samples fixed in formalin, embeded in paraffin and stored for 1 and 5 years in the Patology Department at UNIFESP
Grantee:Edna Sadayo Miazato Iwamura
Support Opportunities: Regular Research Grants