Leptospirosis / Toxoplasma nucleic acid detection real time taqman
probe PCR kit
Intended use:
This kit is suitable for the specific detection of leptospirosis
(Lep) and Toxoplasma (TOX) in EDTA whole blood samples of cats and
dogs.
Principle:
This kit selects the leptospirosis (Lep) and Toxoplasma (TOX)
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, The nucleic acids of
leptospirosis and Toxoplasma gondii were detected by fluorescence
PCR apparatus in a completely closed reaction system.
Main components:
| Component | Specification |
| PCR reaction liquid | 8 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
Feline/dog EDTA whole blood samples
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℃.
Using Micgene 242/244/244IVD and ABI QuantStudio5:
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 and VIC/HEX are selected for detection channels.
The results of interpretation
| FAM channel(Lep) | VIC/HEX channel (TOX) | Result interpretation |
| 1 | - | - | Results were reported negative |
| 2 | + | - | Results were reported Lep positive |
| 3 | - | + | Results were reported TOX positive |
| 4 | + | + | Results were reported Lep,TOX positive |
Note: "+" means positive, "-" means negative
Positive judgment value or reference interval
| Result | Ct Value |
| Positive | Ct≤38 |
| Negative | No Ct value or Ct value=0 |
| 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.
Operation details please check from IFU!
