345 Evaluation and Validation of an Automated Platform for Ribonucleic Acid Extraction From Formalin-Fixed Paraffin-Embedded Tissue

Isolation of high quality ribonucleic acid (RNA) from formalin-fixed paraffin-embedded (FFPE) archival material is often a challenging task due to RNA degradation, which occurs during the formalin fixation process. Presently over 80% of tissue tested in molecular diagnostics laboratories are derived from FFPE samples. Thus, utilizing a robust and reliable RNA extraction protocol is an immensely critical tool for many clinical lab applications. An array of commercially available manual and automated extraction kits on the market promise high-quality RNA yields. Currently our laboratory employs a manual extraction protocol. However, as our clinical operation scales, we decided to pursue an automated RNA extraction protocol that can provide sufficient yield for our clinical assay. Here we compare our FFPE manual RNA extraction protocol, RecoverAll (ThermoFisher, Waltham, MA) with an automated kit, LEV RNA-FFPE (Promega, Madison, WI), to assess whether RNA quality and concentration is adequate and comparable for clinical processing. A search was conducted of our electronic database to identify 12 FFPE cases of varying and ranging tumor quantity and quality that were previously extracted using the manual procedure. To ensure inter- and intra-tech reproducibility, the assay was performed by two technologists in duplicates. RNA quality and quantity was assessed on the Nanodrop (ThermoFisher, Waltham, MA) and Qubit Fluorometer (LifeTechnologies, Carlsbad, CA). At the conclusion of our initial experiment, it was determined that both the manual and automated protocols produced equivalent quantitative and qualitative metrics derived from the Nanodrop and Qubit scores. We determined that the automated protocol offers numerous additional benefits, such as reduced contamination, quicker turnaround time, safer reagent handling, and negligible cost difference. Given its technical advantages and operational benefits, it is apparent the automated protocol is more suitable and reliable for clinical applications. Further studies will be conducted to internally validate according to regulatory standards.