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Guangzhou BioKey Healthy Technology Co.Ltd |
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Multiple Feline Respiratory Pathogens (FCV FHV MF CF) Real -time PCR detection kit (PCR-Fluorescent Probe Method)
Intended use:
This kit is suitable for the specific detection of feline calicivirus (FCV), feline herpes virus (FHV), feline mycoplasma (MF) and feline chlamydia (CF) in cat nasal and oral secretions.
Principle:
This kit uses the conserved gene fragments of feline calicivirus (FCV), feline herpevirus (FHV), feline mycoplasma (MF) and feline chlamydia (CF) to design specific probes, which can specifically bind to a DNA template in the middle of the primer amplification region. During the PCR extension process, The exonuctase activity of Taq enzyme cuts the 5 '-end fluorophore from the probe and causes it to dissociate in the reaction system, thus separating from the 3' -end quenched group and emitting fluorescence, which is detected by fluorescence PCR instrument. The nucleic acids of feline calicivirus (FCV), feline herpes virus (FHV), feline mycoplasma (MF) and feline chlamydia (CF) were detected in the total closed reaction system.
Main components:
| composition | quantity | storage conditions |
| PCR reaction liquid(FCV) | 6 tube | -20±5℃ |
| PCR reaction liquid(FHV) | 6 tube | |
| PCR reaction liquid(MF) | 6 tube | |
| PCR reaction liquid(CF) | 6 tube | |
| Positive control | 1 tube | |
| Negative control | 1 tube |
Note: components in different batches of kits should not be interchangeable.
Sample requirements:
1. Sample type
Cat nasal and mouth secretions, 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℃.
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 | 50℃ | 20min |
| 2 | 1 | 95℃ | 2min30s |
| 3 | 45 | 95℃ | 10s |
| 60℃ | 30s Collect the fluorescent |
FAM was selected for detection channel.
The results of interpretation
| 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 < 40 is considered as
Operation details please check from IFU!
