Applicability of a System for Fully Automated Nucleic Acid Extraction from Formalin-fixed Paraffin-embedded Sections for Routine KRAS Mutation Testing

Annika Lehmann, Christiane Schewe, Guido Hennig, Carsten Denkert, Wilko Weichert, Jan Budczies, Manfred Dietel

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Due to the approval of various new targeted therapies for the treatment of cancer, molecular pathology laboratories with a diagnostic focus have to meet new challenges: simultaneous handling of a large number of samples, small amounts of input material, and fragmentation of nucleic acids because of formalin fixation. As a consequence, fully automated systems for a fast and standardized extraction of high-quality DNA from formalin-fixed paraffin-embedded (FFPE) tissues are urgently needed. In this study, we tested the performance of a fully automated, high-throughput method for the extraction of nucleic acids from FFPE tissues. We investigated the extraction performance in sections of 5 different tissue types often analyzed in routine pathology laboratories (cervix, colon, liver, lymph node, and lung; n=340). Furthermore, we compared the quality, labor input, and applicability of the method for diagnostic purposes with those of a laboratory-validated manual method in a clinical setting by screening a set of 45 colorectal adenocarcinoma for the KRAS mutation. Automated extraction of both DNA and RNA was successful in 339 of 340 FFPE samples representing 5 different tissue types. In comparison with a conventional manual extraction protocol, the method showed an overall agreement of 97.7% (95% confidence interval, 88.2%-99.9%) for the subsequent mutational analysis of the KRAS gene in colorectal cancer samples. The fully automated system is a promising tool for a simple, robust, and rapid extraction of DNA and RNA from formalin-fixed tissue. It ensures a standardization of sample processing and can be applied to clinical FFPE samples in routine pathology.

Original languageEnglish
Pages (from-to)114-119
Number of pages6
JournalDiagnostic Molecular Pathology
Volume21
Issue number2
DOIs
StatePublished - Jun 2012
Externally publishedYes

Keywords

  • FFPE
  • KRAS
  • automated DNA extraction
  • molecular pathology
  • routine diagnostics

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