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V. Harveyi(VH) Detection kit Nucleic Acid Detection Kits
(PCR-Fluorescent Probe Method)
Intended use:
This kit is suitable for the specific detection of pathogenic
Vibrio harveyi (VH) in bait, water samples, shrimp seedlings,
shrimp tissues and processed products.
Principle:
This kit is chosen, V. Harveyi (VH) specific fragment to design
primers probe probe, the probe 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, Vibrio haviovibrio nucleic acid (VH) was detected by
fluorescence PCR instrument in a completely closed reaction system.
Main components:
Component | Specification |
PCR reaction liquid | 24 tubes |
Positive control | 1 tube |
Negative control | 1 tube |
Sample requirements:
1. Sample type:
Shrimp seedling, shrimp tissue, water body, bait, processed
products, 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 -20℃.
Using Micgene 242/244/244IVD and ABI QuantStudio5:
procedures are set as below:
Steps | Cycle number | Temperature | Time |
1 | 1 | 95℃ | 3min |
2 | 45 | 95℃ | 10s |
60℃ | 30sCollect 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 < 45 is considered
as positive, and no Ct value or Ct value ≥45 is considered as
negative.
Result analysis:
Automatic analysis using instrumentation
Quality control:
Negative control: No obvious amplification curve of FAM detection
channel;
Positive control: FAM channel showed obvious amplification curve,
Ct value ≤30;
The above conditions are met simultaneously in the same experiment;
otherwise, the experiment is invalid and re-testing is required.