Abstract
AIMS: Mutation detection accuracy has been described extensively; however, it is surprising that pre-PCR processing of formalin-fixed paraffin-embedded (FFPE) samples has not been systematically assessed in clinical context. We designed a RING trial to (i) investigate pre-PCR variability, (ii) correlate pre-PCR variation with EGFR/BRAF mutation testing accuracy and (iii) investigate causes for observed variation. METHODS: 13 molecular pathology laboratories were recruited. 104 blinded FFPE curls including engineered FFPE curls, cell-negative FFPE curls and control FFPE tissue samples were distributed to participants for pre-PCR processing and mutation detection. Follow-up analysis was performed to assess sample purity, DNA integrity and DNA quantitation. RESULTS: Rate of mutation detection failure was 11.9%. Of these failures, 80% were attributed to pre-PCR error. Significant differences in DNA yields across all samples were seen using analysis of variance (p
Original language | English |
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Pages (from-to) | 111-118 |
Number of pages | 8 |
Journal | Journal of Clinical Pathology |
Volume | 68 |
Issue number | 2 |
Early online date | 27 Nov 2014 |
DOIs | |
Publication status | Published - Feb 2015 |
Keywords
- LUNG CANCER
- MELANOMA
- MOLECULAR PATHOLOGY
- PCR
- diagnostic screening
- Cell Line, Tumor
- DNA Mutational Analysis
- DNA, Neoplasm
- Diagnostic Errors
- Fixatives
- Fluorometry
- Formaldehyde
- Great Britain
- Humans
- Laboratory Proficiency Testing
- Mutation
- Observer Variation
- Paraffin Embedding
- Polymerase Chain Reaction
- Predictive Value of Tests
- Proto-Oncogene Proteins B-raf
- Receptor, Epidermal Growth Factor
- Reproducibility of Results
- Spectrophotometry
- Tissue Fixation
- Transfection
- United States
- Workflow