PCR-Fluorescent probe kit Streptococcus iniae septicaemia virus Nucleic Acid Detection Kit

Intended use:

This kit is suitable for the specific detection of pathogenic Streptococcus iniae septicaemia virus in fish fry or viscera tissue samples such as heart, intestine, spleen and kidney.

Principle:

This kit designed primers with Streptococcus iniae septicaemia virus specific fragment to 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 detection of Streptococcus iniae septicaemia virus in the totally closed reaction system was realized by fluorescence PCR instrument.

Main components:

1. Sample type

Fish fry or fish heart, intestine, spleen, kidney and other visceral tissue tissue samples.

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. Sample preservation

It can be stored for 24 hours at 2℃~8℃, and can be stored for a long time at -20℃.

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 was selected as the detection channel.

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.

The results of interpretation

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!