Diagnostic / Laboratory virus Fluorescence PCR kit Siniperca
chuatsi rhabdovirus Nucleic Acid PCR Kit
Intended use:
This kit is suitable for auxiliary diagnosis of Siniperca chuatsi
rhabdovirus (SCRV) infection in animal tissue, feed, feces and
other samples.
Principle:
This kit to choose a conservative design specific gene probe, the
probe can with primer amplification area in the middle of Siniperca
chuatsi rhabdovirus (SCRV) 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, The detection of Siniperca chuatsi
rhabdovirus (SCRV) nucleic acid in the completely closed reaction
system was realized by fluorescence PCR instrument.
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
Samples of animal tissue, feed, feces, etc
2. Sample collection and transportation
About 0.1g tissue samples were taken from prawn samples according
to different sizes or infection stages. In which, whole individuals
were taken from larvae and larvae, head and chest of eyes were
removed from juveniles under 3CM, gill area was taken from larger
juveniles, and gill filaments or heart or lymphatic organs were
taken from adult shrimps.
3. Storage and transportation
Samples are transported in ice bags of 2℃~8℃, and repeated freezing
and thawing are prohibited. Short-term storage at -20℃, -70℃ can be
long-term storage
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℃ | 5min |
| 2 | 1 | 95℃ | 5min |
| 3 | 45 | 95℃ | 10s |
| 60℃ | 30s collect fluorescent |
FAM was selected for detection channel
The results of interpretation:
| Results | Ct Value |
| Positive | Ct≤40 |
| Negative | No Ct value or Ct value=0 |
| 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.
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 10
copies/μL.
3. Precision: Coefficient of variation (CV,%) of intra-batch
precision Ct value is ≤5%.
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.
Operation details please check from IFU!
