Bacteria DNA Detection Kit (qPCR), BP-QN99

Detects bacterial DNA contamination by probe-based qPCR.

> 92% of bacteria

~60,000 subtypes matched

100 CFU

Limit of detection

20 min

Sample processing, no special equipment

Product overview

Bacteria DNA Detection Kit (qPCR), Cat. No. BP-QN99-50, is a fluorescent-probe-based qPCR kit intended for qualitative detection of bacterial DNA contamination in biological samples. This assay targets conserved bacterial 16S rRNA coding regions and incorporates an internal control for workflow-performance monitoring.

Features and applications

Features

Simple operation: Sample processing requires no special equipment and can be completed in approximately 20 minutes.
Detection types: Matches approximately 92% (about 291,800 of 318,200) of bacterial 16S 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 fungi.
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.

Applications

The kit is used for qualitative detection of bacteria DNA contamination in cells, cell and gene therapy products, and other biological products.

Components

Components	Specification/Notes
Bac qPCR MIX	1.0 mL x 1 tube
Bac Positive Control	1.5 mL x 2 tubes
Bac Negative Control	1.5 mL x 2 tubes
Bac Internal Control	1.0 mL x 1 tube
Instructions	1 copy

Performance data

qPCR Sensitivity Verification

Organism (100 CFU)	Detection rate
Staphylococcus aureus	24/24
Pseudomonas aeruginosa	24/24
Bacillus subtilis	24/24
Clostridium sporogenes	24/24
Micrococcus luteus	24/24
Salmonella enterica	24/24
Escherichia coli	24/24
Cutibacterium acnes	24/24
Streptococcus pyogenes	24/24

Table 1. Verification of sensitivity using bacterial standard strains. S. aureus, P. aeruginosa, B. subtilis, C. sporogenes, M. luteus, S. enterica, E. coli, C. acnes, and S. pyogenes were obtained from the National Institutes for Food and Drug Control (NIFDC), 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
Candida albicans	Negative	Negative	Negative
Aspergillus niger	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 fungi, and Mycoplasma. No cross-reactivity was observed with MDBK, PK15, CHO, HEK293T, C6, CRFK, Sf9, human genomic DNA, C. albicans, A. niger, and M. pneumoniae DNA.

Coverage Analysis

The kit primers and probe were aligned against the SILVA database. Of approximately 318,200 known bacterial 16S sequences, approximately 291,800 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.

Performance data shown here were obtained with the reference materials, extraction workflows, instruments and acceptance criteria used in BIORI’s product-specific validation study. Users should verify performance in their own sample matrix and workflow.

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.