E. coli Residual DNA Detection Kit (qPCR), BP-QN24
Quantifies residual E. coli DNA by probe-based qPCR.
Product overview
E. coli Residual DNA Detection Kit (qPCR), BP-QN24-100, provides quantitative detection of E. coli residual DNA in intermediates, semi-finished products, and finished biological and pharmaceutical products. This fluorescent-probe-based qPCR assay includes an internal-standard and calibrators for residual host-cell DNA testing.
Features and applications
- qPCR probe-based assay targeting conserved E. coli genomic sequences.
- Kit includes an exogenous internal standard for monitoring extraction and amplification performance.
- UDG enzyme is included to help prevent carry-over contamination derived from amplicons.
- Five E. coli calibrators are supplied from 30 fg/μL to 300 pg/μL.
- The COA reports cover precision, accuracy, amplification efficiency, linear correlation, and internal-standard Ct results.
- Quantification of E. coli residual DNA in biopharmaceutical process samples.
- Testing of intermediates, semi-finished products, and finished products.
- Compatible with qPCR workflows on real-time PCR systems listed in the product manual.
- Sample Extraction Recovery Control (ERC) workflows when paired with a suitable nucleic acid extraction or purification kit.
Components
| Component | Specification |
|---|---|
| E. coli qPCR MIX | 1.0 mL × 2 tubes |
| E. coli Internal Standard | 1.0 mL × 1 tube |
| E. coli Negative Control | 1.0 mL × 2 tubes |
| E. coli Calibrator ST1 | 0.5 mL × 1 tube; 30 fg/μL |
| E. coli Calibrator ST2 | 0.5 mL × 1 tube; 300 fg/μL |
| E. coli Calibrator ST3 | 0.5 mL × 1 tube; 3 pg/μL |
| E. coli Calibrator ST4 | 0.5 mL × 1 tube; 30 pg/μL |
| E. coli Calibrator ST5 | 0.5 mL × 1 tube; 300 pg/μL |
Performance data
Performance Data
E. coli Residual DNA Detection Kit (qPCR) Catalog No. BP-QN24-100
A product-specific validation study evaluated analytical performance for E. coli residual DNA. This released version presents data for linear range, limit of quantitation, limit of detection, and recovery.
Key Performance at a Glance
| Performance parameter | Selected result |
|---|---|
| Linear range | 5 fg/μL to 300 pg/μL |
| Limit of Quantitation (LOQ) | 5 fg/μL |
| Limit of Detection (LOD) | 1 fg/μL |
| Acceptance range for recovery | 70%-130% |
Linearity
Observed in this validation study: across the evaluated range of 5 fg/μL to 300 pg/μL, the coefficient of determination (R2) was 0.9996 and the amplification efficiency was 99.264%.
Limit of Quantitation
Observed in this validation study: at 5 fg/μL, the mean concentration was 4.71E+00 fg/μL, with 12.40% CV and -5.70% accuracy bias across 10 replicates.
| Replicate | Concentration value | Mean concentration | CV | Accuracy bias |
|---|---|---|---|---|
| Replicate 1 | 5.72E+00 | 4.71E+00 | 12.40% | -5.70% |
| Replicate 2 | 4.47E+00 | |||
| Replicate 3 | 4.62E+00 | |||
| Replicate 4 | 4.50E+00 | |||
| Replicate 5 | 4.11E+00 | |||
| Replicate 6 | 5.76E+00 | |||
| Replicate 7 | 4.85E+00 | |||
| Replicate 8 | 4.38E+00 | |||
| Replicate 9 | 4.59E+00 | |||
| Replicate 10 | 4.14E+00 |
Limit of Detection
The national E. coli DNA content reference standard (lot 270027-201101) was diluted to 0.5 fg/μL and 1 fg/μL. Twenty-four replicates were tested at each level. Detection rates were 91.67% at 0.5 fg/μL and 100% at 1 fg/μL; therefore, the kit LOD was 1 fg/μL.
Recovery
Five sample levels (0.0281 pg/μL, 0.281 pg/μL, 2.81 pg/μL, 28.1 pg/μL, and 281 pg/μL) were purified with the pretreatment kit and tested with the amplification kit. Observed recoveries ranged from 87.09% to 95.16% (acceptance range 70%-130%).
| Sample | Concentration 1 | Concentration 2 | Concentration 3 | Mean concentration | CV | Target | Recovery |
|---|---|---|---|---|---|---|---|
| 1 | 2.60E+01 | 2.84E+01 | 2.58E+01 | 2.67E+01 | 5.57% | 2.81E+01 | 95.16% |
| 2 | 2.77E+02 | 2.55E+02 | 2.55E+02 | 2.62E+02 | 4.93% | 2.81E+02 | 93.32% |
| 3 | 2.36E+03 | 2.48E+03 | 2.50E+03 | 2.45E+03 | 3.04% | 2.81E+03 | 87.09% |
| 4 | 2.40E+04 | 2.54E+04 | 2.65E+04 | 2.53E+04 | 4.92% | 2.81E+04 | 89.98% |
| 5 | 2.29E+05 | 2.47E+05 | 2.67E+05 | 2.48E+05 | 7.72% | 2.81E+05 | 88.14% |
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | ≤-20 °C, protected from light. Shelf life: 24 months. Avoid more than 10 freeze-thaw cycles. |
| Shipping | Transport with ice pack cold chain packaging and keep the required frozen storage conditions. |
| Handling | Protect reagents from light and avoid unnecessary freeze-thaw cycles. |
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