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Product Description
MicroSHENTEK® Bacteria DNA Detection Kit is used together with the MicroSHENTEK® Fungi & Bacteria DNA Extraction Kit to qualitatively determine bacterial contamination in cells, cell products, vaccines and related matrices. The assay uses real-time PCR (qPCR) and has been validated against USP, EP, JP and Ch.P requirements. The kit covers approximately 92% of known bacterial species (nearly 60,000 species/subspecies sequences) and has a reported detection limit of not more than 35 CFU per reaction. Specificity and coverage have been validated across a variety of matrices, production cells and engineered fungi. For extraction procedures refer to MicroSHENTEK® Fungi & Bacteria DNA Extraction Kit User Guide (Product No. 1504633). Reagents provided are sufficient for 50 reactions.
Technical Specifications
| Parameter | Details |
|---|---|
| Limit of Detection (LOD) | Not more than 35 CFU/reaction |
| Coverage | Approximately 92% of known bacterial species (~60,000 species/subspecies sequences) |
| Validation / Standards | Validated according to pharmacopeial guidance: USP, EP, JP and Ch.P (references include USP<71>, EP<2.6.21>, EP<2.6.7>, EP<2.6.16>, EP<2.6.1>, JP<4.06>, Ch.P<1101>, Ch.P<0234>) |
| Intended use | Qualitative detection of bacterial DNA to assess bacterial contamination in cells, cell products, vaccines and related samples |
| Number of reactions per kit | 50 reactions |
| Reaction volume | 30 μL total reaction volume (10 μL qPCR MIX + 20 μL sample/control) |
| qPCR thermal cycling | Activation: 95 °C 10:00 (1 cycle); Denaturation: 95 °C 00:15; Annealing: 55 °C 00:30; Extension: 72 °C 01:00; 45 cycles. Fluorescence read during extension. |
| Control acceptance thresholds (FAM channel) | NTC: Ct ≥ 39.00 or no amplification; NCS: Ct ≥ 39.00 or no amplification; PC: Ct < 35.00 with proper S-shaped amplification; PCS: Ct < 39.00 with S-shaped amplification |
| Test sample interpretation (FAM channel) | Positive: Ct < 39.00 (at least one well) with S-shaped amplification. Negative: Ct ≥ 39.00 or absence of specific amplification in duplicate runs. If PCS Ct ≥ 39.00, experimental process needs checking or retest may be recommended. |
| Instrument compatibility | Compatible with common real-time PCR systems (examples: SHENTEK-96S Real-time PCR System, Roche LightCycler 480 II) |
Features
- Real-time qPCR detection: Uses probe-based real-time PCR for sensitive and specific detection of bacterial DNA.
- Validated to pharmacopeial guidance: Assay performance validated according to USP, EP, JP and Ch.P standards for sterility / nucleic acid amplification methods.
- High coverage: Designed to detect approximately 92% of known bacterial species (~60,000 sequences), increasing the likelihood of detecting diverse contaminants.
- Low detection limit: Reported detection limit of ≤35 CFU per reaction (LOD validated during method validation).
- Controls included: Includes positive control and procedural controls to support result interpretation and assay QC.
- Compatibility and flexibility: Works with standard PCR plates/strips and is compatible with multiple real-time PCR instruments; workflow supports separate negative/sample/positive areas to minimize contamination.
Applications
- Qualitative detection of bacterial contamination in cell cultures
- Sterility testing for cell products and biologics (vaccines, therapeutics)
- Quality control during production of biological products
- Screening of production cells and engineered fungi for bacterial DNA contamination
- Research applications requiring sensitive detection of bacterial DNA in various matrices
Kit Contents
Bac Positive Control (PC) (Part No. NNA047) 50 μL × 1 tube-20 °C
Bac qPCR Reaction Buffer (Part No. NNB019) 220 μL × 2 tubes-20 °C (protect from light)
Bac Primer & Probe MIX (Part No. NNC098) 50 μL × 2 tubes-20 °C
DNA Dilution Buffer (DDB) (Part No. NND001) 1.5 mL × 2 tubes-20 °C
* Total: 50 reactions | Method: qPCR
Attention
• For Research Use Only (RUO). Not for human diagnostic use.
• Read Material Safety Data Sheets (MSDS) and follow all handling instructions.
• Wear appropriate PPE: protective eyewear, mask, lab coat and gloves.
• Work areas should be segregated into Negative, Sample and Positive areas; use dedicated equipment in each area to prevent contamination.
• Decontaminate work surfaces, pipettes and tubes by UV irradiation (60 minutes) and 75% ethanol and use nucleic acid scavenger as needed.
• Thaw reagents at 2–8 °C or on ice; vortex and briefly spin before use.
• Prepare dilutions of positive control last and perform dilutions in the positive area to avoid contamination.
• Follow specified plate loading order: add and seal NTC and NCS in negative area, add test samples in sample area, then add PCS and PC in positive area.
• If detecting fungi and bacteria in the same run, add UNG enzyme step (25 °C for 10 minutes) and perform two-channel assay (FAM and VIC) as instructed.
• If control criteria are not met (e.g., PCS Ct ≥39.00), re-check experiment and consider retesting.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the limit of detection (LOD) of the kit?
The kit reports a detection limit of not more than 35 CFU per reaction (LOD as validated during method validation).
Q2: Which real-time PCR instruments are supported?
The kit is compatible with common real-time PCR systems. Examples provided: SHENTEK-96S Real-time PCR System and Roche LightCycler 480 II. Other instruments may be used if configured to the specified reaction volume and cycling program.
Q3: How should the kit be stored and what is the shelf life?
Store kit components at -20 °C. Components are stable for up to 24 months; check the label expiration date.
Q4: What are the acceptance criteria for controls?
NTC and NCS: Ct ≥ 39.00 (or no amplification). PC: Ct < 35.00 with effective S-shaped amplification. PCS: Ct < 39.00 with effective S-shaped amplification. If PCS Ct ≥ 39.00, the process should be rechecked.
Research Use Only
✔
Research Use Only (RUO). Assay performance validated according to USP, EP, JP and Ch.P guidance (e.g., USP<71>, EP<2.6.21>, JP<4.06>, Ch.P references). – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.
✔
Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
