Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz

Logo MUG-Forschungsportal

Gewählte Publikation:

SHR Neuro Krebs Kardio Lipid

Bonin, S; Hlubek, F; Benhattar, J; Denkert, C; Dietel, M; Fernandez, PL; Höfler, G; Kothmaier, H; Kruslin, B; Mazzanti, CM; Perren, A; Popper, H; Scarpa, A; Soares, P; Stanta, G; Groenen, PJ.
Multicentre validation study of nucleic acids extraction from FFPE tissues.
Virchows Arch. 2010; 457(3): 309-317. [OPEN ACCESS]
Web of Science PubMed PUBMED Central FullText FullText_MUG Google Scholar

 

Autor/innen der Med Uni Graz:
Hoefler Gerald
Kothmaier Hannelore
Popper Helmuth
Altmetrics:

Dimensions Citations:

Plum Analytics:

Scite (citation analytics):

Number of Figures: 4
| | | |
Abstract:
In most pathology laboratories worldwide, formalin-fixed paraffin embedded (FFPE) samples are the only tissue specimens available for routine diagnostics. Although commercial kits for diagnostic molecular pathology testing are becoming available, most of the current diagnostic tests are laboratory-based assays. Thus, there is a need for standardized procedures in molecular pathology, starting from the extraction of nucleic acids. To evaluate the current methods for extracting nucleic acids from FFPE tissues, 13 European laboratories, participating to the European FP6 program IMPACTS (www.impactsnetwork.eu), isolated nucleic acids from four diagnostic FFPE tissues using their routine methods, followed by quality assessment. The DNA-extraction protocols ranged from homemade protocols to commercial kits. Except for one homemade protocol, the majority gave comparable results in terms of the quality of the extracted DNA measured by the ability to amplify differently sized control gene fragments by PCR. For array-applications or tests that require an accurately determined DNA-input, we recommend using silica based adsorption columns for DNA recovery. For RNA extractions, the best results were obtained using chromatography column based commercial kits, which resulted in the highest quantity and best assayable RNA. Quality testing using RT-PCR gave successful amplification of 200 bp-250 bp PCR products from most tested tissues. Modifications of the proteinase-K digestion time led to better results, even when commercial kits were applied. The results of the study emphasize the need for quality control of the nucleic acid extracts with standardised methods to prevent false negative results and to allow data comparison among different diagnostic laboratories.
Find related publications in this database (using NLM MeSH Indexing)
Formaldehyde -
Humans -
Nucleic Acids - analysis
Paraffin Embedding -
Pathology, Molecular - methods
Reproducibility of Results -
Reverse Transcriptase Polymerase Chain Reaction -
Tissue Fixation -

Find related publications in this database (Keywords)
FFPE
Multicentre study
Molecular analyses standardisation
PCR
DNA
RNA
Isolation
© Med Uni Graz Impressum