Real Time Fluorescence qPCR Detection System Bacterial / Vibrio
Count (Bac/ Vib) PCR test 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 |
Note: components in different batches of kits should not be
interchangeable.
Sample requirements:
1. Sample type
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 picture:
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!
