Edwardsiella piscicida septicaemia virus (EDW.P) Nucleic Acid DNA
Taq polymerase PCR kit
Intended use:
This kit is suitable for the specific detection of pathogenic
Edwardsiella piscicida septicaemia virus (EDW.P) in fish viscera
tissue samples.
Principle:
This kit selection Edwardsiella piscicida septicaemia virus (Edw)
P) specific fragment to design primers probe probe, 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, Fluorescence was
emitted and detected by fluorescence PCR instrument to achieve the
detection of Edwardsiella piscicida septicaemia virus (EDW.P) in
the 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
Fish liver, intestine, stomach 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 | 95℃ | 3min |
| 2 | 45 | 95℃ | 10s |
| 60℃ | 30s collect fluorescent |
FAM is selected for detection channels.
The results of interpretation:
| Results | Ct Value |
| Positive | t≤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.
Operation details please check from IFU!
