| Field | Specification |
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Description
The Lentivirus Physical Titer qPCR Detection Kit is a kit designed to quantitatively detect the physical titer of lentivirus (total number of viral particles) using the conserved region (LTR) in the genome of HIV-1-derived lentiviral vectors as a target.
This kit adopts the principle of fluorescence quantitative PCR by probe method to rapidly and specifically detect lentivirus RNA. The widest linear range can reach 5 copies/μL~5×108 copies/μL, with a detection limit as low as 0.5 copies/μL. And it contains the Lentivirus RNA Control (RNA quantitative reference product). The kit needs to be used together with the the Residual DNA Sample Preparation Kit (Cat# 18461ES) and Recombinant DNase I (RNase-free) (Cat#10325ES).
Features
- Compliance with regulations: the products are fully validated according to the requirements of Chp, USP, ICH, etc., and their performance is in line with Chinese and foreign regulations and standards;
- Guarantee of quality: the raw materials of the kits are all self-developed, and the qPCR Mix and other enzyme products are produced in an ultra-clean enzyme factory;
- High sensitivity: the lower limit of quantification (LLOQ) was 5 copies/uL;
- High precision: high intra-batch reproducibility CV <10%, small inter-batch variation i.e. intermediate precision CV <15%;
- Strong specificity: specific detection of Lentivirus RNA without interference from other exogenous genomic DNA;
- Strong anti-interference: the introduction of internal quality control (IC) can exclude sample interference, abnormal reaction preparation and other factors, effectively avoiding false negatives.
Application
- Residual Lentivirus RNA test in biological products.
Specifications
|
Sensitivity |
5 copies/uL (range 5～5×108copies/μL）) |
|
Assay Time |
~1.5 hours |
|
Assay Principle |
Fluorescent probe qPCR method |
|
Required Instruments |
Real-time fluorescence quantitative PCR system |
Product Performance
|
Product Performance Validation Program |
Reference standards or requirements |
YEASEN |
|
|
1. Standards for the kit |
Homemade standards are available |
Plasmid RNA containing the conserved region (LTR) in the genome of HIV-1-derived lentiviral vectors |
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2. Linear Range |
Assay Range |
Refer to the actual situation |
5~5x108 copies/μL |
|
R2 |
≥0.98 |
≥0.99 and above |
|
|
Slope |
-3.1~-3.8 |
-3.1~-3.6 |
|
|
Amplification Efficiency |
84%~109% |
90%~109% |
|
|
3. Accuracy |
Recovery Rate |
50%~150% |
70%~130% and above |
|
4. Precision |
Repeatability |
CV≤15% |
＜10% |
|
Intermediate precision |
CV≤15% |
＜15% |
|
|
5. Exclusivity |
No interference with RNA from other sources |
Specific detection of Lentivirus RNA without interference from other exogenous RNA |
|
|
6. Sensitivity |
Limit of quantification |
Reliable quantification of the smallest concentrations of target substances |
5 copies/μL |
|
Limit of detection |
Refer to the actual situation |
0.5 copies/μL |
|
|
7. Durability |
Reagent Stability |
Evaluating the effect of different storage temperatures on reagent performance |
37°C for 7 days, and repeated freeze-thaw 10 times, the kit amplification efficiency is in the range of 90%~109%, R2≥0.99 and above |
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Instrument Compatibility |
Kits are used on different devices without affecting the test results |
Compatible with many types of qPCR Instruments (eg. Bio-Rad CFX96 Optic Module, Thermo Scientific ABI 7500, ABI Quant Studio 5) |
|
Components
|
Components No. |
Name |
41333ES50 |
41333ES60 |
|
41333-A |
Lentivirus qPCR Mix |
0.75 mL |
1.5 mL |
|
41333-B |
One Step Enzyme Mix |
200 μL |
400 μL |
|
41333-C |
RNA Dilution Buffer |
1.8 mL×2 |
1.8 mL×4 |
|
41333-D |
Lentivirus RNA Control (5×108 copies/μL) |
25 μL |
50 μL |
|
41333-E |
IC* |
50 μL |
100 μL |
*IC：Internal control.
Storage
This product should be stored at -25~-15℃ for 1 years.
The 41333-A should be stored protected from light.
Figures
- Limit of quantification
Lentivirus RNA at concentrations of 10 copies/uL, 5 copies/uL and 2 copies/uL were detected, with 10 replicates for each concentration, respectively. The results showed that the CV of Lentivirus RNA was <20% at concentrations of 5 copies/μL and above. Therefore, the limit of quantification in the Lentivirus RNA detection kits was 5 copies/μL for Lentivirus RNA.
Table 1. Limit of quantification (LOQ) assay results for Lentivirus RNA
|
Repetition number
Test item |
Lentivirus RNA (copies/μL) |
|
|
Quantity |
Return ratio |
|
|
1 |
5.43 |
108.60% |
|
2 |
5.7 |
114.00% |
|
3 |
4.93 |
98.60% |
|
4 |
5.72 |
114.40% |
|
5 |
4.76 |
95.20% |
|
6 |
5.53 |
110.60% |
|
7 |
6.53 |
130.60% |
|
8 |
4.95 |
99.00% |
|
9 |
4.84 |
96.80% |
|
10 |
5.67 |
113.40% |
|
Average value |
5.41 |
\ |
|
CV |
13.49% |
\ |
Figure 1. 5 copies/μL Lentivirus RNA qPCR assay results
- Limit of detection
The detection of Lentivirus RNA at various concentration points under the quantitative limit, combined with the blank limit requirements, yields the detection limit of the reagent kit. The results show that at concentrations of 0.5 copies/μL and above, in 20 repeated wells, ≥19 wells are detected (i.e., the detection rate is ≥95%). Therefore, the detection limit of the Lentivirus Physical Titer qPCR Detection Kit can reach 0.5 copies/μL.
Table 2. Limit of detection (LOD) assay results for Lentivirus RNA
|
Ct value Theoretical concentration |
Lentivirus RNA (copies/μL) |
||
|
1 |
0.5 |
0.2 |
|
|
Positive count / Total count |
20/20 |
20/20 |
18/20 |
|
Detection rate |
100% |
100% |
90% |
Documents:
Safety Data Sheet
Manuals
41333_Manual_Ver.EN20251219.pdf
Replication-competent virus (RCV) — including RCL (lentivirus), RCR (retrovirus), and rcAAV — can form when recombination events during viral vector production reconstitute a replication-competent genome. Detection is mandatory for gene therapy product release under FDA (21 CFR 312.23), EMA, and ICH Q5A guidelines. Even trace amounts of RCV pose significant biosafety risks in clinical applications due to their potential to integrate and replicate in patient cells.
This qPCR-based kit achieves high sensitivity with an LOD typically in the range of 1–10 copies per reaction for RCL/RCR detection, and less than 10 copies per reaction for rcAAV2 detection. Specificity is confirmed against wild-type viral sequences not present in replication-deficient vector preparations. Refer to the product datasheet for validated LOD, LOQ, and specificity data established with certified reference materials.
This kit provides a research-grade (RUO) molecular tool for RCV detection with sensitivity appropriate for process development and QC screening. For GMP lot release submissions to regulatory agencies, full assay qualification (sensitivity, specificity, linearity, robustness) must be conducted per applicable guidelines (ICH Q2R1, USP <1033>). Consider this kit as a validated starting platform for developing a GMP-qualified method.
This kit is validated for purified nucleic acid extracted from viral vector preparations, cell culture supernatants, and drug substance/product samples. Pre-analytical steps (extraction method, input volume) should be optimized for your specific matrix. For complex matrices such as formulated drug products containing stabilizers or preservatives, spike-recovery experiments should confirm assay performance before routine use.
Yeasen Biotechnology offers flexible customization options for many of its assay kits and detection reagents, including custom lot sizes, bulk ordering, and application-specific formulation adjustments. Volume pricing, custom packaging, and kit bundling may be available depending on the product and intended workflow. A Certificate of Analysis (CoA) and lot-specific QC data are provided with every order. For inquiries regarding large-volume orders, custom configurations, or integration into automated workflows, please contact the BioHippo team for a tailored quotation.
