Multicentre validation study of nucleic acids extraction from FFPE tissues

• DOI: 10.1007/s00428-010-0917-5
• Publication Date: 2010-07-27T10:22:12Z
• Authors: Serena Bonin; Falk Hlubek; Jean Benhattar; Carsten Denkert; Manfred Dietel; Pedro L. Fernandez; Gerald Höfler; Hannelore Kothmaier; Bozo Kruslin; Chiara Maria Mazz...; Aurel Perren; Helmuth Popper; Aldo Scarpa; Paula Soares; Giorgio Stanta; Patricia J. T. A....
• Keywords: Tissue Fixation; FFPE . Multicentre study . Molecular analyses standardisation . PCR . DNA . RNA . Isolation; FFPE; Multicentre study; Molecular analyses standardisation; PCR; DNA; RNA; Isolation; Multicentre study; FFPE; FFPE . Multicentre study . Molecular analyses standardisation . PCR . DNA . RNA . Isolation; Isolation; Pathology and Forensic Medicine; 03 medical and health sciences; 0302 clinical medicine; Formaldehyde; Nucleic Acids; Humans; Pathology, Molecular; FFPE; multicenter study; molecular analyses; Molecular Biology; FFPE; Multicentre study; Molecular analyses standardisation; PCR; DNA; RNA; Isolation; Paraffin Embedding; Reverse Transcriptase Polymerase Chain Reaction; Reproducibility of Results; DNA; Cell Biology; 3. Good health; Formaldehyde; Humans; Nucleic Acids/analysis; Nucleic Acids/isolation & purification; Paraffin Embedding; Pathology, Molecular/methods; Pathology, Molecular/standards; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction; Tissue Fixation; Molecular analyses standardisation; PCR; ONCOL 3: Translational research; RNA; Original Article

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