Applying context of use to quantitative polymerase chain reaction method validation and analysis: a recommendation from the European Bioanalysis Forum

Bioanalysis. 2021 Dec;13(23):1723-1729. doi: 10.4155/bio-2021-0218. Epub 2021 Oct 28.

Abstract

Polymerase chain reaction (PCR) is widely used in various fields of laboratory testing, ranging from forensic, molecular biology, medical and diagnostic applications to a wide array of basic research purposes. COVID-19 infection testing has brought the three-letter PCR abbreviation into the vocabulary of billions of people, making it likely the most well-known laboratory test worldwide. With new modalities and translational medicine gaining importance in pharmaceutical research and development, PCR or more specifically, quantitative PCR (qPCR) is now becoming a standard tool in the (regulated) bioanalytical laboratory, driving the bioanalytical community to define best practices for method development, characterization and validation. In absence of specific guidance from health authorities, qPCR may be vulnerable to scope creep from pharmacokinetics (PK) assay validation as defined in bioanalytical method validation guidance/guidelines. In this manuscript, the European Bioanalysis Forum builds a rationale for applying context of use principles when defining requirements for qPCR assay performance and validation criteria.

Keywords: assay development; assay validation; biomarker; context of use; polymerase chain reaction; quantitative polymerase chain reaction; regulated bioanalysis.

MeSH terms

  • Biological Assay / methods*
  • Europe
  • Humans
  • Polymerase Chain Reaction / methods*
  • Research Design