This poster presents a case study on quantitative PCR (qPCR) assay development and validation for detecting adeno-associated virus (AAV) vector shedding across five biological matrices: whole blood, serum, semen, urine, and saliva. The study supports cell and gene therapy and vaccine clinical trials by evaluating key performance parameters such as limit of detection (LOD), lower limit of quantification (LLOQ), linearity, accuracy, precision, and selectivity.
Key findings include:
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All matrices demonstrated acceptable accuracy and precision, with semen requiring dilution due to higher baseline variability.
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LOD and LLOQ values were generally below 1000 copies/mL, except for semen.
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Excellent linearity was observed across matrices, with regression slopes close to 1.0.
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An investigation into reagent variability revealed that pH acidification of the MagMAX Viral/Pathogen (MVP) bead binding solution over time contributed to significant differences in Ct values, potentially impacting assay sensitivity and clinical trial reliability.
The study emphasizes the importance of matrix-specific optimization and quality control in PCR-based bioanalysis for gene therapy safety monitoring.
