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Best Practices in qPCR and dPCR Validation in Regulated Bioanalytical Laboratories

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A Correction to this article was published on 19 April 2022

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Abstract

The use of molecular techniques in the bioanalytical laboratory is becoming more common as the number of gene and cell therapy products continues to increase. Currently, there is no bioanalytical regulatory guidance for these techniques, and contract research organizations are dependent on scientific judgment and best practice to execute this work to GxP compliant status for preclinical and clinical studies supporting biodistribution and vector shedding. This manuscript describes the process and rationale for development and validation of quantitative polymerase chain reaction (qPCR) and digital PCR (dPCR) assays as presented in a 2021 AAPS two-day workshop on the harmonization of qPCR. The scope, herein, includes bioanalytical validation parameters and acceptance criteria utilizing these technologies. Additionally, pros and cons of these molecular techniques will be highlighted, and the common pitfalls to avoid will be illustrated. The aim of this manuscript is to provide best-practice, working recommendations, and the facilitation of future regulatory guidance.

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Authors and Affiliations

  1. BioAgilytix Labs, 2300 Englert Drive, Durham, NC, 27713, USA

    Amanda Hays & Dave Williams

  2. Smithers, Gaithersburg, MD, USA

    Rafiq Islam

  3. PPD, Richmond, VA, USA

    Katie Matys

Authors
  1. Amanda Hays
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  2. Rafiq Islam
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  3. Katie Matys
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  4. Dave Williams
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Contributions

A.H, R.I, K.M, and D.W contributed equally in writing of this manuscript.

Corresponding author

Correspondence to Amanda Hays.

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Conflict of Interest Statement

The authors are employed by and receive compensation from companies that are involved in development and validation of qPCR methods and are listed on the title page of the manuscript. The authors have no other relevant affiliations or financial involvements with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.

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Hays, A., Islam, R., Matys, K. et al. Best Practices in qPCR and dPCR Validation in Regulated Bioanalytical Laboratories. AAPS J 24, 36 (2022). https://doi.org/10.1208/s12248-022-00686-1

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