Advancing Lentiviral Genome Integrity and Titer Analysis Via Two Dimensional Reverse Transcription-digital PCR (2D RT-dPCR)

Lentiviral vectors (LVVs) are powerful and versatile gene‑transfer tools widely used in both ex vivo and in vivo cell and gene therapies. Traditional single‑channel dPCR or qPCR assay only provides limited information of the viral genome and cannot distinguish between complete and partial genomes. To address these limitations, we developed a lentiviral genome integrity and titer assay using 2D RT‑dPCR. This method enables comprehensive characterization of the LVV genome—including genomic titer and genome integrity—in a single reaction. When combined with traditional infectious titer assays, our method provides deeper insights into LVV product quality and quantity.