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Cat. No.: VRDDK-50 (for 50T)

Cat. No.: VRDDK-200 (for 200T)

Description

Vero Residual DNA Detection Kit (qPCR) is a probe‑based quantitative PCR (qPCR) assay designed for rapid, efficient, and highly sensitive quantification of residual genomic DNA derived from Vero host cells in various biological product samples. This kit incorporates an anti‑contamination system containing high‑quality UDG enzyme and dUTP at optimized ratios, effectively eliminating false‑positive results or abnormally low Ct values caused by carryover contamination from previous PCR amplifications.

Vero cells (African Green Monkey Kidney Cells) are a continuous aneuploid cell line capable of undergoing numerous passages without senescence and are suitable for large‑scale culture. Derived from the kidney epithelial cells of the African green monkey, Vero cells are one of the World Health Organization–recommended substrates for viral propagation in the production of human and veterinary vaccines. They are widely used in vaccine manufacturing and are the preferred cell line for rabies vaccine production. Vero cells are also used for producing poliovirus, measles vaccines, and for industrial‑scale production of recombinant proteins and antibody therapeutics.

Biological products manufactured using Vero cells may still contain trace amounts of Vero cell–derived DNA fragments even after rigorous purification. These fragments may carry infectious or oncogenic risks, and elevated levels of residual DNA have been associated with increased risks of oncogenicity, infectivity, and immune dysregulation. Regulatory agencies in many countries and regions impose strict limits on host cell DNA residues in biopharmaceutical products. Monitoring and controlling host cell DNA levels is essential for evaluating immunogenicity risk and ensuring product safety.

This kit enables highly sensitive detection of residual DNA from Vero host cells, with a limit of detection in the femtogram (fg) range. Based on the quantitative PCR method described for exogenous DNA residue testing, the probe‑based qPCR workflow achieves a detection limit of 3 fg/μl and a quantitative range of 3 fg/μl to 300 pg/μl.

Components

• Probe qPCR Mix (2X, UDG)
• Primer/Probe Mix (10X)
• Ultrapure Water
• Dilution Buffer
• Vero DNA Control (30ng/μl)
• Low ROX (50X)
• High ROX (50X)

Features

• This kit exhibits high specificity, and genomic DNA from CHO cells and E. coli does not interfere with Vero DNA residue detection. The assay exclusively detects Vero host cell DNA and shows no cross‑reactivity with DNA from other species.
• This kit demonstrates excellent stability, and varying degrees of DNA fragmentation have minimal impact on the detection of Vero DNA residues. Vero DNA Control was fragmented to different extents using micrococcal nuclease (MNase), followed by qPCR analysis with this kit. The results showed that DNA fragments longer than 200 bp were consistently detected without affecting assay performance. However, when fragmentation was excessively severe and the majority of DNA fragments were shorter than 200 bp, the measured DNA residue levels tended to be underestimated.
• This kit demonstrates excellent repeatability. When Vero DNA at 3 pg/μl and 30 fg/μl was tested in 12 replicates, the coefficient of variation (CV) for both concentrations was less than 5%.
• This kit provides the essential components required for qPCR, including the Probe qPCR Mix (2X, UDG) and a premixed set of specific primers and probe. The premix contains high‑quality, Vero‑DNA–free Taq DNA polymerase, UDG enzyme, PCR buffer, dNTPs, dUTP, stabilizers, and magnesium ions, covering all universal reaction components. The high‑purity primer–probe mixture is designed using unique tandem repeat sequences within the Vero genome as the detection target, enabling ultra‑sensitive quantitative detection of Vero DNA. In addition, the kit includes a Vero DNA Control, which serves as a positive control to verify kit performance and to generate standard curves.
• This product is available in both Low ROX and High ROX formats, ensuring broad compatibility with real‑time PCR instruments that require no ROX, Low ROX, or High ROX as a passive reference dye. ROX functions as an internal fluorescence normalization dye to correct for non‑PCR‑related signal fluctuations, thereby minimizing well‑to‑well variation. Such variation may arise from factors including pipetting errors or sample evaporation. Different qPCR instruments have different ROX requirements; therefore, users should select the appropriate Probe qPCR Mix (2X) containing High ROX, Low ROX, or no ROX based on the instrument in use. In general, Probe qPCR Mix (2X, High ROX) is also compatible with instruments that require Low ROX or no ROX.