DNA Taq polymeraset detect Grass Carp Reovirus virus ( GCRVI ) Type
I PCR test kit
Intended use:
This kit is suitable for auxiliary diagnosis of Grass Carp Reovirus
Type I virus (GCRVI) infection
Principle:
This kit uses RNA as a template and primers as a starting point to
synthesize cDNA strands complementary to RNA template under the
action of high-efficiency reverse transcriptase. Choose peach virus
gene conservative design specific 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, The nucleic acid
of taora virus was detected by fluorescence PCR in a completely
closed reaction system.
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
Tissue samples of grass carp and black carp
2. Sample collection
Fry with body length ≤4cm should be taken as a whole fish, if there
is yolk sac, it should be removed; All viscera including kidney and
brain were taken from 4~6cm fry. Liver, brain, spleen and kidney
were taken from fish with body length ≥6cm. 5 fish can be combined
into a small sample.
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℃ | 20min |
| 2 | 1 | 95℃ | 2min30s |
| 3 | 45 | 95℃ | 10s |
| 60℃ | 30s collect fluorescent |
FAM is selected for detection channels.
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
3copies/μL.
3. Precision: Coefficient of variation (CV,%) of intra-batch
precision Ct value is ≤5%.
Operation details please check from IFU!
