Mycobacterium DNA Detection Kit (qPCR), BP-QN15
50-reaction qPCR kit for qualitative Mycobacterium DNA contamination detection.
Product overview
This Mycobacterium DNA Detection Kit (qPCR), Cat. No. BP-QN15-50, is a fluorescent-probe-based qPCR kit intended for qualitative detection of Mycobacterium DNA contamination in biological samples. The kit employs specific primers and probes targeting conserved regions of the Mycobacterium genome to amplify Mycobacterium-derived DNA, featuring simple workflow, fast turnaround, high specificity and high analytical sensitivity.
Features and applications
- Simple operation: The workflow from sample extraction through completion of testing takes only 1.5 hours.
- Limit of detection: 100 CFU
- Precision: Intermediate precision (Ct value) CV ≤5%.
- Specificity: No cross-reactivity with common cell lines and structurally related microorganisms.
- The kit is used for qualitative detection of Mycobacterium DNA contamination in cells, cell and gene therapy products, and other biological products.
Components
| Components | Specification/Notes |
|---|---|
| Mycobacterium qPCR MIX | 1.0 mL x 1 tube |
| Mycobacterium Positive Control | 1.5 mL x 2 tubes |
| Mycobacterium Negative Control | 1.5 mL x 2 tubes |
| Mycobacterium Internal Control | 1.0 mL x 1 tube |
| Instructions | 1 copy |
Performance data
qPCR Sensitivity Verification
Table 1. Summary of Limit-of-Detection Data for 12 Common Standard Mycobacterium Strains
| Strain | Positive/total | Strain | Positive/total |
|---|---|---|---|
| Mycobacterium avium (aliquot of national reference standard) | 24/24 | Mycobacterium terrae (aliquot of national reference standard) | 24/24 |
| Mycobacterium shimoidei (aliquot of national reference standard) | 24/24 | Mycobacterium kansasii (aliquot of national reference standard) | 24/24 |
| Mycobacterium asiaticum (aliquot of national reference standard) | 24/24 | Mycobacterium scrofulaceum (aliquot of national reference standard) | 24/24 |
| Mycobacterium gordonae (aliquot of national reference standard) | 24/24 | Mycobacterium chelonae (aliquot of national reference standard) | 24/24 |
| Mycobacterium fortuitum (aliquot of national reference standard) | 24/24 | Mycobacterium phlei (aliquot of national reference standard) | 24/24 |
| Mycobacterium tuberculosis (aliquot of national reference standard) | 24/24 | Mycobacterium smegmatis (ATCC) | 24/24 |
The Mycobacterium DNA Detection Kit detected every standard Mycobacterium strain in 24 of 24 tests.
Cross-Reactivity Results
Cross-reactivity was evaluated using structurally related microorganisms and engineered cell lines. DNA from 29 panel-listed items (Lactobacillus acidophilus, Propionibacterium, Escherichia coli, Enterobacter aerogenes, Acinetobacter baumannii, Salmonella enterica, Pseudomonas aeruginosa, Campylobacter jejuni, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus intermedius, Micrococcus luteus, Streptococcus agalactiae, Lactococcus lactis, Streptococcus pneumoniae, Candida albicans, Streptococcus mutans, Pichia pastoris, Monascus purpureus, Trichoderma harzianum, SP2/0, Vero, HEK293T, CHO, BHK21, NS0, Sf9, and MDCK) showed no cross-reactivity with the Mycobacterium detection reagent. The results are shown in Table 2.
Table 2. Cross-Reactivity Results
| No. | Sample | Test result | No. | Sample | Test result |
|---|---|---|---|---|---|
| 1 | L. acidophilus | Negative | 16 | S. pneumoniae | Negative |
| 2 | Propionibacterium | Negative | 17 | C. albicans | Negative |
| 3 | E. coli | Negative | 18 | S. mutans | Negative |
| 4 | E. aerogenes | Negative | 19 | P. pastoris | Negative |
| 5 | A. baumannii | Negative | 20 | M. purpureus | Negative |
| 6 | S. enterica | Negative | 21 | T. harzianum | Negative |
| 7 | P. aeruginosa | Negative | 22 | SP2/0 | Negative |
| 8 | C. jejuni | Negative | 23 | Vero | Negative |
| 9 | S. aureus | Negative | 24 | HEK293T | Negative |
| 10 | S. epidermidis | Negative | 25 | CHO | Negative |
| 11 | S. saprophyticus | Negative | 26 | BHK21 | Negative |
| 12 | S. intermedius | Negative | 27 | NS0 | Negative |
| 13 | M. luteus | Negative | 28 | Sf9 | Negative |
| 14 | S. agalactiae | Negative | 29 | MDCK | Negative |
| 15 | L. lactis | Negative | 30 | Negative control | Negative |
Conclusion: DNA from the 29 panel-listed items (L. acidophilus, Propionibacterium, E. coli, E. aerogenes, A. baumannii, S. enterica, P. aeruginosa, C. jejuni, S. aureus, S. epidermidis, S. saprophyticus, S. intermedius, M. luteus, S. agalactiae, L. lactis, S. pneumoniae, C. albicans, S. mutans, P. pastoris, M. purpureus, T. harzianum, SP2/0, Vero, HEK293T, CHO, BHK21, NS0, Sf9, and MDCK) showed no cross-reactivity with the Mycobacterium detection reagent.
Precision
Within-Run Precision
In-house reference materials Z1, Z2, and Z3 were tested in 10 replicates each. Precision CV was within 5% at every concentration.
Intermediate Precision
Intermediate precision was evaluated across operators, days, instruments, and reagent lots using in-house reference materials Z1, Z2, and Z3, with five replicates per material. Precision CV was within 5% at every concentration.
Total Imprecision
Data across operators, days, instruments, and reagent lots were pooled. Precision CV was within 5% at every concentration.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | ≤-20°C, protected from light. Shelf life: 24 months. Avoid more than 7 freeze-thaw cycles. |
| Shipping | Ship frozen using dry-ice or validated frozen cold-chain packaging to maintain required frozen-storage conditions. |
| Handling | Protect from light and avoid unnecessary freeze-thaw cycles where applicable. |
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