Fast Amplification Viral Haemorrhagic Septicemia Virus VHSV PCR Test Kit

Fast Amplification PCR kit Viral haemorrhagic Septicemia Virus ( VHSV ) test kit ( fluorescent Probe Method )

 
Intended use:

This kit is suitable for auxiliary diagnosis of Viral haemorrhagic Septicemia Virus (VHSV) infection in fish fry or viscera tissue samples such as fish heart, intestine, spleen and kidney.

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 viral haemorrhagic septicemia virus gene conservative design specific 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, Fluorescence is emitted and detected by fluorescence PCR instrument to realize nucleic acid detection of viral haemorrhagic septicemia virus in the completely closed reaction system.
 
Main components:
 

1. Sample type

Fish fry or viscera tissue samples such as heart, intestine, spleen, kidney, etc.

2. Sample collection and transportation

Samples shall be collected and transported in accordance with SC/T 7103. Samples shall be transported to the laboratory as soon as possible under refrigerated conditions to avoid repeated freeze-thaw.

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:

FAM is selected for detection channels.

Quality control:

1. Negative control: No obvious amplification curve of FAM detection channel;
2. Positive control: FAM channel showed obvious amplification curve, Ct value ≤30;
3. The above conditions are met simultaneously in the same experiment; otherwise, the experiment is invalid and re-testing is required.

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 10copies/μL.

3. Precision: Coefficient of variation (CV,%) of intra-batch precision Ct value is ≤5%.

4. Linear, linear correlation coefficient | r | > 0.980.

Product picture:

  Operation details please check from IFU!