Fungi DNA Detection Kit (qPCR), BP-QN78
Detects fungal DNA contamination by probe-based qPCR.
Product overview
Fungi DNA Detection Kit (qPCR), Cat. No. BP-QN78-50, is a fluorescent-probe-based qPCR kit intended for qualitative detection of fungal DNA contamination in biological samples. This assay targets conserved fungal 18S rRNA coding regions and incorporates an internal control for workflow-performance monitoring.
Features and applications
- Simple operation: Sample processing requires no special equipment and can be completed in approximately 20 minutes.
- Detection types: Matches approximately 92% (8,485 of 9,258) of fungal 18S sequences in the SILVA database.
- Limit of detection: 100 CFU
- Precision: Intermediate precision (Ct value) CV ≤5%.
- Specificity: No cross-reactivity with common cell lines and bacteria.
- Internal control: Internal control (IC) can be added during the sample extraction phase to evaluate extraction efficiency, or incorporated during the PCR amplification reaction phase to monitor potential inhibition of the amplification process and prevent false-negative results.
- Contamination control: This kit contains UDG enzyme to prevent contamination of amplification products.
- Applicable devices: Including but not limited to SLAN-96P and SLAN-96S fully automated medical PCR analysis systems; ABI7500 and ABI QuantStudio™ 5 real-time fluorescent quantitative PCR systems; Roche LightCycler 480 fluorescent quantitative PCR system; and Bio-Rad CFX96 quantitative PCR system.
- The kit is used for qualitative detection of fungi DNA contamination in cells, cell and gene-therapy products, and other biological products.
Components
| Components | Specification/Notes |
|---|---|
| Fun qPCR MIX | 1.0 mL x 1 tube |
| Fun Positive Control | 1.5 mL x 2 tubes |
| Fun Negative Control | 1.5 mL x 2 tubes |
| Fun Internal Control | 1.0 mL x 1 tube |
| Instructions | 1 copy |
Performance data
qPCR Sensitivity Verification
| Organism (100 CFU) | Detection rate |
|---|---|
| Candida albicans | 24/24 |
| Aspergillus niger | 24/24 |
Table 1. Verification of sensitivity using fungal reference strains. Candida albicans and Aspergillus niger were obtained from the National Institutes for Food and Drug Control (NIFDC) and China Medical Culture Collection (CMCC).
Cross-Reactivity Results
| Sample | Replicate 1 | Replicate 2 | Replicate 3 |
|---|---|---|---|
| MDBK | Negative | Negative | Negative |
| PK15 | Negative | Negative | Negative |
| CHO | Negative | Negative | Negative |
| HEK293T | Negative | Negative | Negative |
| C6 | Negative | Negative | Negative |
| CRFK | Negative | Negative | Negative |
| Sf9 | Negative | Negative | Negative |
| human genomic DNA | Negative | Negative | Negative |
| Pseudomonas aeruginosa | Negative | Negative | Negative |
| Staphylococcus aureus | Negative | Negative | Negative |
| Escherichia coli | Negative | Negative | Negative |
| Mycoplasma pneumoniae | Negative | Negative | Negative |
Table 2. Cross-Reactivity Results The kit was evaluated against DNA from commonly used cell lines, common bacteria, and Mycoplasma. No cross-reactivity was observed with MDBK, PK15, CHO, HEK293T, C6, CRFK, Sf9, human genomic DNA, P. aeruginosa, S. aureus, E. coli, and M. pneumoniae DNA.
Coverage Analysis
The kit primers and probe were aligned against the SILVA database. Of 9,258 fungal 18S sequences in the database, 8,485 matched, corresponding to approximately 92% sequence coverage.
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 | ≤-18 °C, protected from light. Shelf life: 24 months. Avoid more than 7 freeze-thaw cycles. |
| Shipping | Ship frozen on dry ice or with validated frozen cold-chain packaging that maintains the specified frozen storage condition. |
| Handling | Protect from light and avoid unnecessary freeze-thaw cycles where applicable. |
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