Product Design and Validation References:
- Chinese Pharmacopoeia 2020 Edition: 1101 Sterility Test Method
- Chinese Pharmacopoeia 2020 Edition: 9201 Pharmaceutical Microbiological Test Alternative Method Validation Guidelines
- “Human Stem Cell-derived Pharmaceutical Research and Evaluation Technical Guidelines (Trial)”
- “Immune Cell-derived Pharmaceutical Research and Evaluation Technical Guidelines (Trial)”
- American Pharmacopoeia USP44: Sterility Tests
- Japanese Pharmacopoeia JP17: Sterility Tests
- Polymerase Chain Reaction (PCR) Laboratory Inspection Guidelines (2016 Edition)
Product Name | Catalog No. | Product Specification | Storage Conditions |
Bacteria Detection Kit (qPCR Method) | BA0001-50 | The test kit contains 5×BAC reaction solution, enzyme, BAC positive control, BAC internal control, PCR dilution buffer, and 50 tests/kit. | Transport and store at -20℃ Shelf life: 12 months. |
Details
- This product is used for the rapid qualitative detection of bacterial contamination in culture media, cell culture materials, and biological products, and test results can be obtained in 2 hours.
- It can detect five types of bacteria used for sterility testing in the pharmacopoeia: Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans, and Escherichia coli. The detection limit is not more than 100 CFU/mL. The detection limits for Pseudomonas aeruginosa and Bacillus subtilis are verified to be 0.5 CFU/mL. The database comparison covers nearly 400 types of bacteria.
- Using FAM and ROX two-color fluorescent probes, it can detect bacterial target sequences and internal quality control respectively. It has good specificity and can eliminate false-negative results caused by PCR-inhibiting factors in samples. There is no cross-reaction with fungi, mycoplasma, NK cells, and 293 cells and other human derived cells.
Performance Verification I: Specificity
It has been verified that it can detect five kinds of reference bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella, and Escherichia coli) mentioned in the Chinese Pharmacopoeia at a concentration of 100 CFU/mL, and has no cross-reaction with fungi, mycoplasmas, 293 cell DNA, NK cell DNA, and human umbilical cord mesenchymal stem cell DNA.
Test Kit Specificity Verification Results (Detected Bacterial Channel Amplification Ct Values) | |||||||
Bacteria | Results | Fungi | Results | Mycoplasmas | Results | Cells | Results |
Bacillus subtilis | 32.04 | Aspergillus niger | NoCt/NoCt | Mycoplasma hyorhinis | NoCt/NoCt | 293 cells | NoCt/NoCt |
Salmonella | 25.46 | Candida albicans | NoCt/NoCt | Mycoplasma orale | NoCt/NoCt | NK cells | NoCt/NoCt |
Escherichia coli | 30.46 | Candida tropicalis | NoCt/NoCt | Mycoplasma pneumoniae | NoCt/NoCt | Stem cells | NoCt/NoCt |
Pseudomonas aeruginosa | 34.10 | Candida glabrata | NoCt/NoCt | Unidentified mycoplasma | NoCt/NoCt | ||
Staphylococcus aureus | 35.92 | ||||||
Performance Verification II: Detection Limit and Sensitivity
Taking Pseudomonas aeruginosa as an example, using a bacterial liquid concentration of 10-0.05 CFU/mL, the results show that the detection limit of Pseudomonas aeruginosa is 0.5 CFU/mL.
Taking Salmonella as an example, using a bacterial liquid concentration of 5-0.05 CFU/mL, the results show that the detection limit of Salmonella is 0.5 CFU/mL.
Performance Verification III: Repeatability and Stability Testing
This test kit was repeatedly produced for 3 batches to test the five types of reference bacteria at a concentration of 100 CFU/mL after extraction. The results are shown in the table below. It can stably detect the five types of reference bacteria at 100 CFU/mL, and the results of the three batches are consistent. The test kit has good reproducibility, with a CV < 10%.
Verification Results of the Repeatability and Stability of the Test Kit (Amplification Ct Values) | ||||||
Batch 1 | Batch 2 | Batch 3 | ||||
Bacteria | Bacteria | Internal Control | Bacteria | Internal Control | Bacteria | Internal Control |
Bacillus subtilis | 32.04 | 30.04 | 35.19 | 29.53 | 35.91 | 31.79 |
Salmonella | 25.46 | 29.59 | 28.01 | 29.13 | 26.32 | 33.80 |
Escherichia coli | 30.46 | 31.10 | 30.59 | 29.56 | 30.28 | 31.63 |
Pseudomonas aeruginosa | 34.10 | 32.00 | 33.72 | 29.75 | 32.11 | 31.11 |
Staphylococcus aureus | 35.92 | 31.39 | 40.55 | 29.40 | 38.65 | 30.59 |
NTC | NoCt | NoCt | NoCt | NoCt | NoCt | NoCt |
NTC | NoCt | NoCt | NoCt | NoCt | NoCt | NoCt |
*For other related experimental validations, please refer to the “Technical White Paper” of Yocon Biotechnology’s products.