Infectious Myonecrosis Virus Nucleic Acid Detection Kits
(PCR-Fluorescent Probe Method)
Intended use:
This kit is suitable for the diagnosis of Infectious Myonecrosis
Virus (IMNV).
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
Edible prawn, machijiming prawn, vannamei and other susceptible
prawn tissues
2. Sample collection
About 0.1g tissue samples were taken from prawn samples according
to different sizes or infection stages. Among them, 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 shrimp.
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.
Using Micgene 242/244/244IVD and ABI QuantStudio5:
procedures are set as below:
| Step | Cycle number | Temperature | Time |
| 1 | 1 | 50℃ | 20min |
| 2 | 1 | 95℃ | 2min30s |
| 3 | 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=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.
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.
Operation details please check from IFU!
