Recombinant Adeno-associated virus (AAV) based gene delivery makes possible transformative and curative treatment options for genetic disorders and is also the leading platform in the field. Studying shedding profiles of viral particles is important to understand the potential risk associated with transmission of the virus and associated gene therapy to individuals (for example, friends, family members, and healthcare workers) and the environment. AAV may be transmitted through respiratory, gastrointestinal, sexual, and possibly vertical (mother to fetus, although pregnant women are currently excluded from gene therapy trials) routes. Therefore, levels of AAV in a comprehensive set of challenging matrices such as whole blood, serum, semen, urine, stool, and buccal swabs are routinely monitored to understand the risks holistically. Given the challenges of detection in the outlined matrices, optimizing the sensitivity of detection can provide important safety data to support clinical development. Currently, quantitative polymerase chain reaction (qPCR) assays are considered the gold standard for studying viral shedding. We will describe the development and validation of a sensitive, reproducible, and robust client specific AAV qPCR assay for use in viral shedding studies.

Kristen Mickey is a Research Scientist III at Eurofins Viracor Biopharma Services where she established herself as a valuable member of the Molecular and Live Cell R&D teams. She has over 9 years of industry experience and has taken the lead and excelled on numerous PCR development and validation projects. Kristen has also developed assays on other platforms, including flow cytometry and viral infectivity assays. She has worked previously as a clinical laboratory scientist and at the Stowers Institute for Medical Research. Kristen has a Master’s in Toxicology from the University of Kansas Medical Center.