Aquatic channel catfish septicaemia virus ( CCV ) DNA taq polymerase RT qPCR detection
Intended use:
This kit is suitable for Channel catfish virus septicaemia virus (CCV) of liver, spleen and kidney tissue samples.
Principle:
This kit selection Channel catfish virus septicaemia virus (CCV) 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, glow, Fluorescence PCR instrument was used to detect Channel catfish virus septicaemia virus CCV 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
Channel catfish”Liver, spleen, kidney, etc2.
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.
2. 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 fluorescent |
FAM is selected for detection channels.
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.
Operation details please check from IFU!
Product Tags:
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Catfish Septicaemia Virus Aquaculture Test Kit CCV RT QPCR Detection DNA Taq Images
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