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Multicentre validation study of nucleic acids extraction from FFPE tissues

The application of molecular tests has become feasible in formalin-fixed paraffin embedded (FFPE) samples. At the present time most of the methods applied in molecular pathology are laboratory-based assays and commercial kits not directly intended for diagnostic purposes. Therefore there is a need to establish guidelines with standardised procedures for molecular methods, basically starting from the extraction of nucleic acids. To evaluate the current methods for extracting nucleic acids from FFPE tissues, 13 different European laboratories, members of the IMPACTS group, isolated nucleic acids from different types of human tissues. Our results clearly show that the quantity and quality of extracts are tissue and method dependent for both nucleic acids. The quality of DNA was not highly affected by the use of different protocols. However, for applications requiring standardized DNA-concentration, silica based adsorption columns for purifying DNA are recommended. For RNA extractions, tissue quality and extraction methods are more critical. The overall best results were obtained by chromatography column based methods, which resulted in highest quantity and quality RNA. Home made methods and commercial monophasic solutions are anyway suitable for many applications. The presented results emphasize the need for quality control of the nucleic acid extracts with standardised methods to avoid false negative results that are unacceptable in diagnostic assays. This comprehensive study may contribute to establish standardised procedures for future high quality molecular diagnostics.

FFPE; multicenter study; molecular analyses

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