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Guangzhou BioKey Healthy Technology Co.Ltd |
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Bacterial/Vibrio Count (Bac/Vib) Nucleic Acid Medical Laboratory Diagnostic PCR Kit
Intended use:
This kit is used for real-time detection of bacteria and vibrio by fluorescence probe PCR method, and is suitable for rapid quantitative detection of bacteria and vibrio by FAM fluorescence channel and HEX fluorescence channel.
Principle:
This kit to choose total bacteria/vibrio specific fragment probes
the design of primers, the probe can with primer amplification area
in the middle of a specific binding DNA template, in the process of
PCR extension, the circumscribed Taq polymerase enzymes from the 5
'end of fluorescent groups on the probe will be cut down, make it
free in the reaction system, thereby quenching groups out of the 3'
end, The total number of bacteria/vibrio nucleic acid in the total
closed reaction system can be detected by fluorescence PCR
instrument.
Main components:
| Component | Specification |
| PCR reaction liquid | 24 tubes |
| Positive control | 1 tube |
| Negative control | 1 tube |
Aquaculture water, prawns, etc.
2. Transport
Sample collection and transportation shall be carried out in accordance with NY/T 541. Samples shall be transported to the laboratory as soon as possible under refrigerated conditions to avoid repeated freeze-thaw.
3. Sample preservation
It can be stored for 24 hours at 2℃~8℃, and can be stored for a long time at -20℃.
PCR amplification and fluorescence detection (PCR detection area):
Put each reaction tube into the PCR instrument and perform PCR amplification according to the following procedures:
| Step | Cycle number | Temperature | Time |
| 1 | 1 | 55℃ | 5min |
| 2 | 1 | 94℃ | 5min |
| 3 | 40 | 94℃ | 10s |
| 60℃ | 25s collect fluorescent |
FAM and HEX/VIC were selected for detection channels
Product performance index:
1. Accuracy
The positive reference materials of testing enterprises were all positive.
2. Minimum detection limit: the detection limit of this kit is 10copies/μL.
3. Precision: Coefficient of variation (CV,%) of intra-batch precision Ct value is ≤5%.
Quality control:
(1) Quality control of total bacterial count detection in FAM fluorescence channel:
Negative control: CT value should be ≥29, positive control: CT value should be ≤25.
(2)HEX/VIC fluorescent channel vibrio count detection quality control:
Negative control: CT value should be ≥35, positive control: CT value should be ≤30.
Operation details please check from IFU!
