Generic Protocols for the Analytical Validation of Next-Generation Sequencing-Based ctDNA Assays: A Joint Consensus Recommendation of the BloodPAC's Analytical Variables Working Group.

Liquid biopsy, particularly the analysis of circulating tumor DNA (ctDNA), has demonstrated considerable promise for numerous clinical intended uses. Successful validation and commercialization of novel ctDNA tests have the potential to improve the outcomes of patients with cancer. The goal of the Blood Profiling Atlas Consortium (BloodPAC) is to accelerate the development and validation of liquid biopsy assays that will be introduced into the clinic. To accomplish this goal, the BloodPAC conducts research in the following areas: Data Collection and Analysis within the BloodPAC Data Commons; Preanalytical Variables; Analytical Variables; Patient Context Variables; and Reimbursement. In this document, the BloodPAC's Analytical Variables Working Group (AV WG) attempts to define a set of generic analytical validation protocols tailored for ctDNA-based Next-Generation Sequencing (NGS) assays. Analytical validation of ctDNA assays poses several unique challenges that primarily arise from the fact that very few tumor-derived DNA molecules may be present in circulation relative to the amount of nontumor-derived cell-free DNA (cfDNA). These challenges include the exquisite level of sensitivity and specificity needed to detect ctDNA, the potential for false negatives in detecting these rare molecules, and the increased reliance on contrived samples to attain sufficient ctDNA for analytical validation. By addressing these unique challenges, the BloodPAC hopes to expedite sponsors' presubmission discussions with the Food and Drug Administration (FDA) with the protocols presented herein. By sharing best practices with the broader community, this work may also save the time and capacity of FDA reviewers through increased efficiency.

Molecular and cellular approaches to patient management in oncology.

Many of the techniques that are employed today by pathologists and oncologists to generate a diagnosis, prognosis or prediction of response have not changed over several decades. However, new molecular and cellular technologies will enable more precise and objective decision-making. This review will detail some of the more recent developments in these areas from Veridex, LLC, academic laboratories and other commercial entities. The discussion of molecular technologies will focus on breast sentinel lymph node biopsy, prostate biopsy, carcinoma of unknown primary, prediction of recurrence in lymph node negative colon and breast cancers and pharmacogenomics. The discussion of cellular technologies will focus on the use of circulating cells to serve in both prognostic and predictive capacities and on the use of molecular methods to interrogate the DNA and RNA isolated from these circulating cells.