Pre-PCR processing of FFPE samples leads to discrepancies in BRAF and EGFR mutation detection
Variation in pre-PCR processing of FFPE samples leads to discrepancies in BRAF and EGFR mutation detection: a diagnostic RING trial
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.
In this study, we provide evidence demonstrating that variation in pre-PCR steps is prevalent and may detrimentally affect the patient’s ability to receive critical therapy. We provide recommendations for preanalytical workflow optimisation that may reduce errors in down-stream sequencing and for next-generation sequencing library generation.
Joshua R Kapp, Tim Diss, James Spicer, Michael Gandy, Iris Schrijver, Lawrence J Jennings, Marilyn M Li, Gregory J Tsongalis, David Gonzalez de Castro,
Julia A Bridge, Andrew Wallace, Joshua L Deignan, Sandra Hing, Rachel Butler, Eldo Verghese, Gary J Latham, Rifat A Hamoudi
Kapp JR, et al. J Clin Pathol 2015;68:111–118. doi:10.1136/jclinpath-2014-202644