The Biospecimen Preanalytical Variables Program: A Multiassay Comparison of Effects of Delay to Fixation and Fixation Duration on Nucleic Acid Quality

Context.— Despite widespread use of formalin-fixed, paraffin-embedded (FFPE) tissue in clinical and research settings, potential effects of variable tissue processing remain largely unknown. Objective.— To elucidate molecular effects associated with clinically relevant preanalytical variability, the National Cancer Institute initiated the BiospecimenPreanalytical Variables (BPV) program. Design.— The BPV program, a well-controlled series of systematic, blind and randomized studies, investigated whether a delay to fixation (DTF) or time in fixative (TIF) affects the quantity and quality of DNA and RNA isolated from FFPE colon, kidney, and ovarian tumorsin comparison to case-matched snap-frozen controls. Results.— DNA and RNA yields were comparable among FFPE biospecimenssubjected to different DTF and TIF time points. DNA and RNA quality metrics revealed assay- and time point–specific effects of DTF and TIF. A quantitative reverse transcription–polymerase chain reaction(qRT-PCR) assay was superior when ...