Nocardia seriolae Nucleic Acid Detection Kit (Real time
PCR-Fluorescent Probe method)
Intended use:
The kit is used for the epidemiological analysis of Nocardia
seriolae in fish viscera tissue samples
Principle:
In this kit, the specific fragment of N.seriolae is used to design
the primer probe probe, which can specifically bind to a SECTION of
DNA template in the middle of the primer amplification region.
During the PCR extension process, the exonuclide activity of Taq
enzyme cuts the 5 '-terminal fluorophore from the probe and makes
it free in the reaction system. Therefore, the quenched 3 '-end
group was separated and the fluorescence was emitted. The
fluorescence PCR instrument was used to detect Nocardia seriolae in
a completely closed reaction system
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
Gills, spleen, kidney and other pathological tissues.
2. Sample collection and transportation
Samples shall be collected and transported in accordance with SC/T
7103. 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 | 94℃ | 3min |
| 2 | 45 | 94℃ | 10s |
| 60℃ | 20s collect fluorescent |
FAM was selected as the detection channel.
Quality control:
- Negative control: No obvious amplification curve of FAM detection
channel;
- Positive control: FAM channel showed obvious amplification curve,
Ct value ≤30;
- The above conditions are met simultaneously in the same experiment;
otherwise, the experiment is invalid and re-testing is required.
The results of interpretation
| Results | Ct Value |
| Positive | Ct≤38 |
| Negative | No Ct value or Ct value≥45 |
| Gray area | 38＜Ct<45 |
Note: The result is judged to be grey area, which needs to be
rechecked. If the recheck result is Ct value < 45 is considered
as positive, and no Ct value or Ct value ≥45 is considered as
negative.
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%.
4. Linear, linear correlation coefficient | r | > 0.980.
Product picture:
Operation details please check from IFU!
