Hot topic discussions have been ongoing with members of the newly formed qPCR Working Group, under auspices of the AAPS Bioanalytical community, including best practices on establishing sensitivity for qPCR assays in regulated bioanalysis. Industry experts have diverse methods for defining the sensitivity of qPCR assays by statistical or empirical approaches that are not harmonized across the industry. In addition, it remains unclear on applicability of the 50 copies per microgram of DNA for the LLOQ in guidance documents to all qPCR assays. In this Hot Topic presentation, we propose a 20-30 minute presentation to introduce the topic and summarize the collected survey results on establishing sensitivity. The presentation will be followed by a 30-35 minute expert panel discussion, to include a regulator if possible, to discuss the survey results, best practices, input on approaches for determining sensitivity and how to apply regulatory guidelines related tp qPCR sensitivity.
Learning Objectives:
Upon completion, participants will be able to understand the basic approaches experts are using to determining sensitivity for qPCR assays.
Upon completion, participants will be able to appreciate the advantages and disadvantages of empirical and statistical approaches to calculating sensitivity for qPCR assays.
Upon completion, participants will be able to realize the diverse uses of LOD/LLOQ for qPCR assays and opportunities and limitations for applying targeted sensitivity of 50 copies per microgram DNA.
