Ehrlichia Canis ( E.C ) Nucleic Acid amplification PCR testing
Fluorescent taqman Probe kit
Intended use:
This kit is suitable for the specific detection of Ehrlichia canis
(E.C) in Canine EDTA whole blood samples.
Principle:
This kit use Ehrlichia canis conservative design specific gene
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 was used to detect Ehrlichia canis nucleic
acid in a completely closed reaction system.
Main components:
Component | Quantity | Storage condition |
| PCR reaction liquid | 8 tubes | -20±5℃ |
| Positive control | 1 tube |
| Negative control | 1 tube |
Note: components in different batches of kits should not be
interchangeable.
Sample requirements:
1. Sample type
Canine EDTA whole blood etc.
2. Sample collection and transportation
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 -70℃.
Method calibration:
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.
Positive judgment value or reference interval
| Results | Ct Value |
| Positive | Ct≤40 |
| Negative | No Ct value or Ct value≥45 |
| Gray area | 40＜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.
Operation details please check from IFU!
