Description
Lentivirus (LV) is a widely used viral vector in cell and gene therapies. However, rare recombination events within the plasmid sequences used to generate LVs can lead to the formation of replication-competent lentivirus (RCL), which can infect non-target cells and replicate, posing significant safety risks. To mitigate these risks, regulatory authorities mandate rigorous quality control testing to ensure the absence of RCLs throughout the lentiviral manufacturing workflow. 1 Accurate detection of RCL is essential for the safety and efficacy of LV-based therapies. Digital PCR, using tools like the QIAcuity RCL Quant Kit, provides unmatched sensitivity and accuracy compared to other methods, such as qPCR, making it an ideal choice for detecting RCLs.
QIAcuity RCL Quant Kit provides: Accurate results on the absence or presence of RCL based on the detection of envelope gene sequences VSV-G DNA (vesicular stomatitis virus G glycoprotein) High sensitivity down to 0.35 copies/µL (Dynamic range of 0.35–7,500 copies/µL) High linearity even in the presence of up to 10 µg gDNA without the need for prior dilution Reproducible results and simplified dPCR workflow
The principle of the dPCR reaction in the nanoplates is described here. QIAcuity RCL Quant Kit detects the absence or presence of RCL based on the detection of envelope gene sequences VSV-G DNA (vesicular stomatitis virus G glycoprotein). It also works in conjunction with dedicated QIAcuity Cell and Gene Therapy dPCR Assays for lentivirus workflow, QIAcuity Digital PCR System and QIAcuity Nanoplates, offering an end-to-end fast dPCR workflow comparable to qPCR but delivering robust detection of RCL in your samples.
QIAcuity RCL Quant Kit is provided for 96 reactions and optimized for RCL detection using the VSV-G Assay and an Internal Control with the QIAcuity ® MasterMix in multiplex reactions on the QIAcuity digital PCR (dPCR) instrument. The VSV-G Assay targets are detected in the Green Channel, while the Internal Control is detected in the Yellow Channel. Both assays come in a ready-to-use primer–probe mix. The QIAcuity RCL Quant Kit is compatible with PvuII restriction enzyme.
